Compositions useful as inhibitors of ERK

ABSTRACT

This invention provides a method of treating a disease state in mammals that is alleviated by treatment with a protein kinase inhibitor, especially an ERK inhibitor, which method comprises administering a compound having the pharmacophoric features Grp1, Grp2 and Grp3:  
                 
 
     Grp 1 is an optionally substituted aryl or aliphatic group; Grp 2 is a heteroaromatic ring having one to three nitrogens, said ring comprising a hydrogen bond acceptor HBA2 optionally bonded to a hydrogen bond donor HBD2, and Grp3 is a heteroaromatic ring comprising a hydrogen bond donor HBD1, with distances between the pharmacophoric features defined in the specification. The method is useful for treating cancer, stroke, diabetes, hepatomegaly, cardiovascular disease, Alzheimer&#39;s disease, cystic fibrosis, viral disease, autoimmune diseases, atherosclerosis, restenosis, psoriasis, allergic reactions, inflammation, neurological disorders or a hormone-related disease.

[0001] This application claims priority to co-pending InternationalPatent Application PCT/US01/03904, filed Feb. 5, 2001, which claimspriority of U.S. Provisional Application serial No. 60/180,502 filedFeb. 5, 2000 and U.S. Provisional Application serial No. 60/191,959filed Mar. 24, 2000. The entirety of which is herein incorporated byreference.

FIELD OF THE INVENTION

[0002] The present invention is in the field of medicinal chemistry andrelates to compounds that are protein kinase inhibitors, compositionscontaining such compounds and methods of use. More particularly, thecompounds are inhibitors of ERK and are useful for treating diseasestates, such as cancer, that are alleviated by ERK inhibitors.

BACKGROUND OF THE INVENTION

[0003] Mammalian mitogen-activated protein (MAP)1 kinases areserine/threonine kinases that mediate intracellular signal transductionpathways (Cobb and Goldsmith, 1995, J Biol. Chem. 270, 14843; Davis,1995, Mol. Reprod. Dev. 42, 459). Members of the MAP kinase family sharesequence similarity and conserved structural domains, and include theERK (extracellular signal regulated kinase), JNK (Jun N-terminal kinase)and p38 kinases. JNKs and p38 kinases are activated in response to thepro-inflammatory cytokines TNF-alpha and interleukin-1, and by cellularstress such as heat shock, hyperosmolarity, ultraviolet radiation,lipopolysaccharides and inhibitors of protein synthesis (Derijard etal., 1994, Cell 76, 1025; Han et al., 1994, Science 265, 808; Raingeaudet al., 1995, J Biol. Chem. 270, 7420; Shapiro and Dinarello, 1995,Proc. Natl. Acad. Sci. USA 92, 12230). In contrast, ERKs are activatedby mitogens and growth factors (Bokemeyer et al. 1996, Kidney Int. 49,1187).

[0004] ERK2 is a widely distributed protein kinase that achieves maximumactivity when both Thr183 and Tyr185 are phosphorylated by the upstreamMAP kinase kinase, MEK1 (Anderson et al., 1990, Nature 343, 651; Crewset al., 1992, Science 258, 478). Upon activation, ERK2 phosphorylatesmany regulatory proteins, including the protein kinases Rsk90 (Bjorbaeket al., 1995, J. Biol. Chem. 270, 18848) and MAPKAP2 (Rouse et al.,1994, Cell 78, 1027), and transcription factors such as ATF2 (Raingeaudet al., 1996, Mol. Cell Biol. 16, 1247), Elk-1 (Raingeaud et al. 1996),c-Fos (Chen et al., 1993 Proc. Natl. Acad. Sci. USA 90, 10952), andc-Myc (Oliver et al., 1995, Proc. Soc. Exp. Biol. Med. 210, 162). ERK2is also a downstream target of the Ras/Raf dependent pathways (Moodie etal., 1993, Science 260, 1658) and may help relay the signals from thesepotentially oncogenic proteins. ERK2 has been shown to play a role inthe negative growth control of breast cancer cells (Frey and Mulder,1997, Cancer Res. 57, 628) and hyperexpression of ERK2 in human breastcancer has been reported (Sivaraman et al., 1997, J Clin. Invest. 99,1478). Activated ERK2 has also been implicated in the proliferation ofendothelin-stimulated airway smooth muscle cells, suggesting a role forthis kinase in asthma (Whelchel et al., 1997, Am. J. Respir. Cell Mol.Biol. 16, 589).

[0005] Based on these findings, ERK signalling has been implicatedvarious disease states including, but not limited to, cancer,inflammation, cardiovascular disease, and neurological disorders, amongothers.

[0006] There is a high unmet medical need to develop protein kinaseinhibitors, especially ERK inhibitors, that are useful in treating thevarious conditions associated with ERK activation, especiallyconsidering the currently available, relatively inadequate treatmentoptions for the majority of these conditions.

[0007] Accordingly, there is still a great need to develop potentinhibitors of protein kinase, including ERK inhibitors, that are usefulin treating various conditions associated with ERK activation.

DESCRIPTION OF THE INVENTION

[0008] It has now been discovered that compounds possessing certainpharmacophoric features as described below are inhibitors of proteinkinase, particularly inhibitors of ERK enzyme. The compounds are usefulin a method for treating a disease state in mammals that is alleviatedby treatment with a protein kinase inhibitor. The method comprisesadministering to a patient in need of such a treatment a therapeuticallyeffective amount of a compound comprising Grp1, Grp2, and Grp3:

[0009] wherein:

[0010] Grp 1 is an optionally substituted aryl or aliphatic group;

[0011] Grp 2 is a heteroaromatic ring comprising one to three nitrogens,and a hydrogen bond acceptor HBA2, wherein HBA2 is optionally bonded toa hydrogen bond donor HBD2; and

[0012] Grp3 is a heteroaromatic ring comprising a hydrogen bond donorHBD1; wherein

[0013] said compound optionally comprises a hydrogen bond acceptor HBA1;and wherein

[0014] Grp1 is within about 2.5-10.0 Å of Grp2; Grp2 is within about4.0-8.0 Å of Grp3; Grp3 is within about 5.0-12.0 Å of Grp1; HBA2 iswithin about 6.5-11.0 Å of Grp1; HBD1 is within about 6.5-8.5 Å of Grp2;HBD1 is within about 3.5-5.5 Å of HBA1; and HBA1 is within about6.7-14.0 Å of HBD2.

[0015] As used herein, the following definitions shall apply unlessotherwise indicated. The term “aliphatic” as used herein means straightchained, branched or cyclic C₁-C₁₂ hydrocarbons which are completelysaturated or which contain one or more units of unsaturation. Forexample, suitable aliphatic groups include substituted or unsubstitutedlinear, branched or cyclic alkyl, alkenyl, alkynyl groups and hybridsthereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or(cycloalkyl)alkenyl. The term “alkyl” and “alkoxy” used alone or as partof a larger moiety refers to both straight and branched chainscontaining one to twelve carbon atoms. The terms “alkenyl” and “alkynyl”used alone or as part of a larger moiety shall include both straight andbranched chains containing two to twelve carbon atoms. The terms“haloalkyl”, “haloalkenyl” and “haloalkoxy” means alkyl, alkenyl oralkoxy, as the case may be, substituted with one or more halogen atoms.The term “halogen” means F, Cl, Br, or I. The term “heteroatom” means N,O or S and shall include any oxidized form of nitrogen and sulfur, andthe quaternized form of any basic nitrogen.

[0016] The term “aryl”, used alone or as part of a larger moiety as in“aralkyl”, refers to aromatic ring groups having five to fourteenmembers, such as phenyl, benzyl, 1-naphthyl, 2-naphthyl, 1-anthracyl and2-anthracyl, and heterocyclic aromatic groups or heteroaryl groups suchas 2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl,5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxadiazolyl,5-oxadiazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 2-pyrrolyl,3-pyrrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl,5-pyrimidyl, 3-pyridazinyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl,5-tetrazolyl, 2-triazolyl, 5-triazolyl, 2-thienyl, or 3-thienyl. Theterm “aryl ring” also refers to rings that are optionally substituted.

[0017] Aryl groups also include fused polycyclic aromatic ring systemsin which a carbocyclic aromatic ring or heteroaryl ring is fused to oneor more other rings. Examples include tetrahydronaphthyl,benzimidazolyl, benzothienyl, benzofuranyl, indolyl, quinolinyl,benzothiazolyl, benzooxazolyl, benzimidazolyl, isoquinolinyl,isoindolyl, acridinyl, benzoisoxazolyl, and the like. Also includedwithin the scope of the term “aryl”, as it is used herein, is a group inwhich one or more carbocyclic aromatic rings and/or heteroaryl rings arefused to a cycloalkyl or non-aromatic heterocyclic ring, for example,indanyl or tetrahydrobenzopyranyl.

[0018] Non-aromatic heterocyclic rings are non-aromatic carbocyclicrings which include one or more heteroatoms such as nitrogen, oxygen orsulfur in the ring. The ring can be five, six, seven or eight-memberedand/or fused to another ring, such as a cycloalkyl or aromatic ring.Examples include 3-1H-benzimidazol-2-one, 3-1-alkyl-benzimidazol-2-one,2-tetrahydrofuranyl, 3-tetrahydrofuranyl, 2-tetrahydrothiophenyl,3-tetrahydrothiophenyl, 2-morpholino, 3-morpholino, 4-morpholino,2-thiomorpholino, 3-thiomorpholino, 4-thiomorpholino, 1-pyrrolidinyl,2-pyrrolidinyl, 3-pyrrolidinyl, 1-piperazinyl, 2-piperazinyl,1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl,4-thiazolidinyl, diazolonyl, N-substituted diazolonyl, 1-phthalimidinyl,benzoxane, benzotriazol-1-yl, benzopyrrolidine, benzopiperidine,benzoxolane, benzothiolane, and benzothiane. The term “heterocyclicring”, whether saturated or unsaturated, also refers to rings that areoptionally substituted.

[0019] An aryl group (carbocyclic and heterocyclic) or an aralkyl group,such as benzyl or phenethyl, may contain one or more substituents.Examples of suitable substituents on the unsaturated carbon atom of anaryl group include a halogen, —R, —OR, —OH, —SH, —SR, protected OH (suchas acyloxy), phenyl (Ph), substituted Ph, —OPh, substituted —OPh, —NO₂,—CN, —NH₂, —NHR, —N(R)₂, —NHCOR, —NHCONHR, —NHCON(R)₂, —NRCOR, —NHCO₂R,—CO₂R, —CO₂H, —COR, —CONHR, —CON(R)₂, —S(O)₂R, —SONH₂, —S(O)R, —SO₂NHR,or —NHS(O)₂R, where R is an aliphatic group or a substituted aliphaticgroup.

[0020] An aliphatic group or a non-aromatic heterocyclic ring maycontain one or more substituents. Examples of suitable substituents onthe saturated carbon of an aliphatic group or of a non-aromaticheterocyclic ring include those listed above for the unsaturated carbonas well as the following: ═O, ═S, ═NNHR, ═NNR₂, ═N—OR, ═NNHCOR,═NNHCO₂R, ═NNHSO₂R, or ═NR.

[0021] The term “alkylidene chain” refers to an optionally substituted,straight or branched, carbon chain that may be fully saturated or haveone or more units of unsaturation. The optional substituents are asdescribed above for an aliphatic group.

[0022] A substitutable nitrogen on an aromatic or non-aromaticheterocyclic ring may be optionally substituted. Suitable substituentson the nitrogen include R, COR, S(O)₂R, and CO₂R, where R is analiphatic group or a substituted aliphatic group.

[0023] The term “linker group” means an organic moiety that connects twoparts of a compound. Linkers are typically comprised of an atom such asoxygen or sulfur, a unit such as —NH— or —CH₂—, or a chain of atoms,such as an alkylidene chain. The molecular mass of a linker is typicallyin the range of about 14 to 200. Examples of linkers include a saturatedor unsaturated C₁₋₆ alkylidene chain which is optionally substituted,and wherein up to two saturated carbons of the chain are optionallyreplaced by —C(═O)—, —CONH—, CONHNH—, —CO₂—, —NHCO₂—, —O—, —NHCONH—,—OC(═O)—, —OC(═O)NH—, —NHNH—, —NHCO—, —O—, —S—, —SO—, —SO₂—, —NH—,—SO₂NH—, or NHSO₂—.

[0024] The terms “backbone chain” and “backbone” refer to the portion ofa polypeptide which comprises the repeating unit —CO—CH—NH—.

[0025] The term “substitute” refers to the replacement of a hydrogenatom in a compound with a substituent group. The term “substitute” doesnot include those hydrogen atoms which form a part of a hydrogen bondingmoiety which is capable of forming a hydrogen bond with a suitablehydrogen bond acceptor, such as a carbonyl oxygen, of an amino acidresidue in the kinase binding site.

[0026] The term “strain energy” is used in this application to refer tothe difference between the conformational energy of the unbound (orfree) compound and that of the compound when bound to kinase. The strainenergy can be determined by the following steps: Evaluate the energy ofthe molecule when it has the conformation necessary for binding tokinase. Then minimize and reevaluate the energy—this is the strainenergy. A more comprehensive definition of strain energy can be found inBostrom, J., Norrby, P.-O.; Liljefors, T., “Conformational EnergyPenalties of Protein-Bound Ligands”, J. Comput. Aided Mol. Design, 1998,383. The strain energy for binding of a potential inhibitor to kinase isthe difference between the free conformation energy and the boundconformation energy. In a preferred embodiment, the strain energy of aninhibitor of the present invention is less than about 10 kcal/mol.

[0027] The term “hydrophobic” refers to a moiety which tends not todissolve in water and is fat-soluble. Hydrophobic moieties include, butare not limited to, hydrocarbons, such as alkanes, alkenes, alkynes,cycloalkanes, cycloalkenes, cycloalkynes and aromatic compounds, such asaryls, certain saturated and unsaturated heterocycles and moieties thatare substantially similar to the side chains of hydrophobic natural andunnatural α-amino acids, including valine, leucine, isoleucine,methionine, phenylanine, α-amino isobutyric acid, alloisoleucine,tyrosine, and tryptophan.

[0028] The term “moderately hydrophobic” refers to a hydrophobic moietyin which one or two carbon atoms have been replaced with one or morepolar atoms, such as oxygen or nitrogen.

[0029] The term “hydrogen bond” refers to a favorable interaction thatoccurs whenever a suitable donor atom, X, bearing a proton, H, and asuitable acceptor atom, Y, have a separation of between 2.5 Å and 3.5 Åand where the angle X-H - - - Y is greater than 90 degrees. Suitabledonor and acceptor atoms are well understood in medicinal chemistry (G.C. Pimentel and A. L. McClellan, The Hydrogen Bond, Freeman, SanFrancisco, 1960; R. Taylor and O. Kennard, “Hydrogen Bond Geometry inOrganic Crystals”, Accounts of Chemical Research, 17, pp. 320-326(1984)).

[0030] The compounds of this invention were designed to bind to proteinkinase, particularly ERK, at the ATP binding site of the enzyme. Thestructure of this binding site has been described for rat ERK2 (BoultonT. G. et al., 1991, Cell 65, 663) and for human ERK2 Owaki H. et al.,1992, Biochem. Biophys. Res. Commun., 182, 1416). The accession numberfor the human ERK2 protein structure in the Swiss-Prot database isP28482.

[0031] The practitioner skilled in the art will appreciate that thereare a number of means to design the inhibitors of the present invention.These same means may be used to select a candidate compound forscreening as an ERK inhibitor. This design or selection may begin withselection of the various moieties which fill binding pockets.

[0032] There are a number of ways to select moieties to fill individualbinding pockets. These include visual inspection of a physical model orcomputer model of the active site and manual docking of models ofselected moieties into various binding pockets. Modeling software thatis well known and available in the art may be used. These include QUANTA[Molecular Simulations, Inc., Burlington, Mass., 1992], SYBYL [MolecularModeling Software, Tripos Associates, Inc., St. Louis, Mo., 1992]. Thismodeling step may be followed by energy minimization with standardmolecular mechanics forcefields such as CHARMM and AMBER. [AMBER: (S. J.Weiner, P. A. Kollman, D. A. Case, U. C. Singh, C. Ghio, G. Alagona, andP. Weiner, J. Am. Chem. Soc., 1984, 106, 765); CHARMM: (B. R. Brooks, R.E. Bruccoleri, B. D. Olafson, D. J. States, S Swaminathan, and M.Karplus, J. Comp. Chem. 1983, 4, 187). In addition, there are a numberof more specialized computer programs to assist in the process ofoptimally placing either complete molecules or molecular fragments intothe protein binding site. These include:

[0033] 1. GRID (Goodford, P. J. A Computational Procedure forDetermining Energetically Favorable Binding Sites on BiologicallyImportant Macromolecules. J. Med. Chem. 1985, 28, 849-857). GRID isavailable from Oxford University, Oxford, UK.

[0034] 2. MCSS (Miranker, A.; Karplus, M. Functionality Maps of BindingSites: A Multiple Copy Simultaneous Search Method. Proteins: Structure,Function and Genetics 1991, 11, 29-34). MCSS is available from MolecularSimulations, Burlington, Mass.

[0035] 3. DOCK (Kuntz, I. D.; Blaney, J. M.; Oatley, S. J.; Langridge,R.; Ferrin, T. E. A Geometric Approach to Macromolecule-LigandInteractions. J. Mol. Biol. 1982, 161, 269-288). DOCK is available fromthe University of California, San Francisco, Calif.

[0036] Once suitable binding orientations have been selected, completemolecules can be chosen for biological evaluation. In the case ofmolecular fragments, they can be assembled into a single inhibitor. Thisassembly may be accomplished by connecting the various moieties to acentral scaffold. The assembly process may, for example, be done byvisual inspection followed by manual model building, again usingsoftware such as Quanta or Sybyl. A number of other programs may also beused to help select ways to connect the various moieties. These include:

[0037] 1. CAVEAT (Bartlett, P. A.; Shea, G. T.; Telfer, S. J.; Waterman,S. CAVEAT: A Program to Facilitate the Structure-Derived Design ofBiologically Active Molecules. In “Molecular Recognition in Chemical andBiological Problems,” Special Pub., Royal Chem. Soc. 1989, 78, 182-196).CAVEAT is available from the University of California, Berkeley, Calif.

[0038] 2. 3D Database systems such as MACCS-3D (MDL Information Systems,San Leandro, Calif.). This area has been recently reviewed by Martin(Martin, Y. C. 3D Database Searching in Drug Design. J. Med. Chem. 1992,35, 2145).

[0039] 3. HOOK (available from Molecular Simulations, Burlington, Mass.)

[0040] In addition to the above computer assisted modeling of inhibitorcompounds, the inhibitors of this invention may be constructed “de novo”using either an empty active site or optionally including some portionsof a known inhibitor. Such methods are well known in the art. Theyinclude, for example:

[0041] 1. LUDI (Bohm, H. J. The Computer Program LUDI: A New Method forthe De Novo Design of Enzyme Inhibitors. J. Comp. Aid. Molec. Design.1992, 6, 61-78). LUDI is available from Biosym Technologies, San Diego,Calif.

[0042] 2. LEGEND (Nishibata, Y., Itai, A., Tetrahedron, 1991, 47, 8985).LEGEND is available from Molecular Simultations, Burlington, Mass.

[0043] 3. LeapFrog (available from Tripos Associates, St. Louis, Mo.)

[0044] A number of techniques commonly used for modeling drugs may beemployed (For a review, see: Cohen, N. C.; Blaney, J. M.; Humblet, C.;Gund, P.; Barry, D. C., “Molecular Modeling Software and Methods forMedicinal Chemistry”, J. Med. Chem., 1990, 33, 883). There are likewisea number of examples in the chemical literature of techniques that canbe applied to specific drug design projects. For a review, see: Navia,M. A. and Murcko, M. A., Current Opinions in Structural Biology, 1992,2, 202. Some examples of these specific applications include: Baldwin,J. J. et al., J. Med. Chem., 1989, 32, 2510; Appelt, K. et al., J. Med.Chem., 1991, 34, 1925; and Ealick, S. E. et al., Proc. Nat. Acad. Sci.USA, 1991, 88, 11540.

[0045] Using the novel combination of steps of the present invention,the skilled artisan can advantageously reduce time consuming andexpensive experimentation to determine enzymatic inhibition activity ofparticular compounds. The method also is useful to facilitate rationaldesign of kinase inhibitors and therapeutic or prophylactic treatmentsagainst kinase-mediated diseases. Accordingly, the present inventionrelates to such inhibitors.

[0046] A variety of conventional techniques may be used to carry outeach of the above evaluations as well as the evaluations necessary inscreening a candidate compound for protein kinase inhibiting activity.Generally, these techniques involve determining the location and bindingproximity of a given moiety, the occupied space of a bound inhibitor,the amount of complementary contact surface between the inhibitor andprotein, the deformation energy of binding of a given compound and someestimate of hydrogen bonding strength and/or electrostatic interactionenergies. Examples of conventional techniques useful in the aboveevaluations include: quantum mechanics, molecular mechanics, moleculardynamics, Monte Carlo sampling, systematic searches and distancegeometry methods [G. R. Marshall, Ann. Rev. Pharmacol. Toxicol., 1987,27, 193]. Specific computer software has been developed for use incarrying out these methods. Examples of programs designed for such usesinclude: Gaussian 92, revision E.2 [M. J. Frisch, Gaussian, Inc.,Pittsburgh, Pa. ©1993]; AMBER, version 4.0 [P. A. Kollman, University ofCalifornia at San Francisco, ©1993]; QUANTA/CHARMM [MolecularSimulations, Inc., Burlington, Mass. ©1992]; and Insight II/Discover[Biosysm Technologies Inc., San Diego, Calif. ©1992]. These programs maybe implemented, for instance, using a Silicon Graphics Indigo2workstation or personal computer network. Other hardware systems andsoftware packages will be known and of evident applicability to thoseskilled in the art.

[0047] Different classes of active protein kinase inhibitors, accordingto this invention, may interact in similar ways with the various bindingregions of the protein kinase active site, particularly the ERK activesite. The spatial arrangement of these important groups is oftenreferred to as a pharmacophore. The concept of the pharmacophore hasbeen well described in the literature [D. Mayer, C. B. Naylor, I. Motoc,and G. R. Marshall, J. Comp. Aided Molec. Design, 1987, 1, 3; A.Hopfinger and B. J. Burke, in Concepts and Applications of MolecularSimilarity, 1990, M. A. Johnson and G. M. Maggiora, Ed., Wiley].

[0048] Different classes of kinase inhibitors of this invention may alsouse different scaffolds or core structures, but all of these cores willallow the necessary moieties to be placed in the active site such thatthe specific interactions necessary for binding may be obtained. Thesecompounds are best defined in terms of their ability to match thepharmacophore, i.e., their structural identity relative to the shape andproperties of the active site of the kinase enzyme such as ERK.

[0049] Distances to or from any given group are calculated from thecenter of mass of that group. The term “center of mass” refers to apoint in three-dimensional space which represents a weighted averageposition of the masses that make up an object. Distances between groupsmay be readily determined using any pharmacophore modeling software andother suitable chemical structure software. Examples of pharmacophoremodeling software that are commercially available include:

[0050] 1. DISCO (Martin, Y. C., Bures, M. G., Danaher, E. A., DeLazzer,J., Lico, A., Pavlik, P. A., J. Comput. Aided Mol. Design, 1993, 7, 83).DISCO is available from Tripos Associates, St. Louis, Mo.

[0051] 2. CHEM-X which is developed and distributed by Chemical DesignLtd, Oxon, UK and Mahwah, N.J.

[0052] 3. APEX-3D which is part of the Insight molecular modelingprogram, distributed by Molecular Simulations, Inc., San Diego, Calif.

[0053] 4. CATALYST (Sprague, P. W., Perspectives in Drug Discovery andDesign, 1995, 3, 1; Müller, K., Ed., ESCOM, Leiden) CATALYST isdistributed by Molecular Simulations, Inc., San Diego, Calif.

[0054] A typical hydrogen bond acceptor (HBA) is an oxygen or nitrogen,especially an oxygen or nitrogen that is sp²-hybridized or an etheroxygen. A typical hydrogen bond donor (HBD) is an oxygen, nitrogen, orheteroaromatic carbon that bears a hydrogen. When bound to ERK,compounds of this invention bind to the ATP binding site of the enzyme.During such binding, Grp1 will occupy a region of the ATP binding sitebounded (within about 5.0 Å of Grp1) by the following amino acids:Val39, Thr110, Asp111, Lys114, and Leu 156. The amino acid numberingused herein corresponds to that of the human ERK2 enzyme. This aminoacid numbering corresponds to the Swiss-Prot database entry foraccession #P28482. The Swiss-Prot database is an international proteinsequence database distributed by the European Bioinformatics Institute(EBI) in Geneva, Switzerland. The database can be found atwww.ebi.ac.uk/swissprot.

[0055] Examples of suitable Grp1 moieties include optionally substitutedaliphatic groups and aryl rings. A preferred Grp1 is an optionallysubstituted phenyl ring.

[0056] Grp2 will occupy a region of the ATP binding site bounded (withinabout 5.0 Å) by the following amino acids: Val39, Ala52, Ile84, Asp106,Leu107, Met108, and Leu156. This relatively small binding pocket favorsmoieties that have a hydrogen bond acceptor HBA2 in a ring where thepositions ortho to the hydrogen bond acceptor have a substituent nolarger than a C₁-C₆ alkyl group. Preferably the positions ortho to HBA₂are substituted with either hydrogen, methyl, or NHR, preferablyhydrogen. Ideally, Grp2 will be capable of orienting in the bindingpocket such that HBA2 can form a hydrogen bond with a backbone aminohydrogen of Met108. In this orientation, hydrogen bond donor HBD2, ifpresent, will form a hydrogen bond with either the backbone carbonyl ofMet108 or the backbone carbonyl of Asp106. Examples of suitable Grp2moieties are listed in Table 1 below. In the examples, the exocycliclines indicate positions where Grp2 may be attached to either Grp1 orGrp3. The distance between Grp2 and Grp1 will be in the range of about2.5 to about 10.0 Å, preferably between about 3.9 to about 8.0 Å, andmost preferably between about 5.7 and 6.8 Å. The distance between HBA2and Grp1 will be in the range of 6.5 to 11.0 Å. The distance betweenGrp2 and Grp3 will be in the range of about 4.0 to about 8.0 Å,preferably between about 5.5 to about 6.6 Å. TABLE 1 Examples of Grp2Moieties

[0057] Grp3 will occupy a region of the ATP binding site bounded by thefollowing amino acids: Val39, Ala52, Lys54, Ile84, Gln105, Asp106,Leu156, and Cys166. One of the positions ortho to the hydrogen bonddonor HBD1 will be either unsubstituted or substituted with a relativelysmall group such as an aliphatic group, mono- or dialkylamino, alkoxy,or thioalkyl having one to four carbons or a halogen such as chlorine.Ideally, Grp3 will be capable of orienting in the binding pocket suchthat HBD1 can form a hydrogen bond with the sidechain carbonyl ofGln105. Examples of suitable Grp3 moieties are shown in Table 2. Thedistance between Grp3 and Grp1 is in the range of about 5.0 to 12.0 Å,preferably between about 6.0 to 10.0 Å, and most preferably betweenabout 7.2 to 8.2 Å. The distance between Grp3 and Grp2 is in the rangeof about 4.0 to 8.0 Å, preferably between about 5.5 to 6.6 Å. Thedistance between HBD1 and Grp1 is in the range of about 7.5 to 11.0 Å,preferably between about 9.0 and 10.0 Å. The distance between HBD1 andGrp2 is in the range of about 6.5 to 8.5 Å, preferably between about 7.2and 8.2 Å. TABLE 2 Examples of Grp3 Moieties

[0058] Compounds of this invention may optionally comprise an additionalhydrogen bond acceptor HBA1 which is closer in distance to Grp3 than itis to Grps 1 or 2. When present, HBA1 will orient in the binding pocketso that it is capable of forming a hydrogen bond with a sidechain aminohydrogen of Lys54, or forming a water bridge to the sidechain carboxylof Glu71, or forming a water bridge to a backbone amino hydrogen ofAsp167. The HBA1 moiety is typically an sp2-hybridized oxygen such asthe carbonyl of a ketone, amide or ester, or the sulfonyl of a sulfone,sulfonamide or sulfate ester. The distance between HBA1 and HBD1 is inthe range of about 3.5 to 5.5 Å, preferably between about 3.9 to 4.9 Å.The distance between HBA1 and HBD2 is in the range of about 7.7 to 14.0Å. When hydrogen bonding to Asp106, the range is between about 7.7 to11.7 Å, preferably between about 10.2 to 11.2 Å. When hydrogen bondingto Met108, the range is between about 11.6 to 13.6 Å, preferably betweenabout 12.1 to 13.1 Å.

[0059] Each of the groups will be connected to at least one other groupby a suitable attachment means such as a valence bond, a suitable linkergroup or by a ring fusion. Suitable linker groups include an alkylidenechain, an aliphatic or aryl ring, —S—, —O—, —CONH—, —SO₂NH—, —NHCO—,—CO—, —NH—, or —NHSO₂—, or a combination thereof.

[0060] One process for designing a kinase inhibitor, particularly an ERKinhibitor, that embodies the present invention comprises the followingsteps. First, one selects a Grp2 moiety that contains a hydrogen bondacceptor HBA2 such as those shown in Table 1. Ideally, the moiety ischosen such that HBA2 would be capable of forming a direct hydrogen bondwith the backbone amino hydrogen of Met108. Second, one selects a Grp1group and a means of attachment to Grp2. It is desirable to confirm thatGrp1 so attached is within the requisite distances to Grp2 and HBA2, andis capable of forming satisfactory interactions with its kinase bindingsite environment as described above. Confirming that the satisfactoryinteractions would be achievable is within the knowledge of one skilledin the art using computational methods such as those described above. Ina like manner, one may build the rest of the inhibitor by selectingGrp3, and optionally HBA1, and corresponding means of attachment toprovide the desired distances between groups and satisfactoryinteractions. The details of utilizing the method for designing an ERKinhibitor of this invention are set forth in the examples.

[0061] The compounds of this invention will usually have a molecularweight of less than about 1000 Daltons, preferably less than about 700Daltons, and more preferably between about 400 and 600 Daltons.

[0062] It will be appreciated that Grp 1, Grp2, and Grp3 may beconnected in various ways while satisfying the requisite distancesdescribed above. For example, one embodiment of this invention relatesto the use of compounds wherein Grps 1 and 3 are each attached to Grp 2as shown in structure type A below, where the dotted lines indicate anattachment means such as a valence bond. Other connectivity schemes arerepresented by structure types B and C:

[0063] The pharmacophoric features of this invention have been used todesign novel kinase inhibitors of structure type A having the generalformula I:

[0064] wherein:

[0065] Ht is a heterocyclic ring selected from pyrrol-3-yl,pyrazol-3-yl, [1,2,4]triazol-3-yl, [1,2,3]triazol-4-yl, ortetrazol-5-yl; said pyrrol-3-yl and pyrazol-3-yl having R³ and QR⁴substituents, and said [1,2,4]triazol-3-yl and [1,2,3]triazol-4-ylsubstituted by either R³ or QR⁴;

[0066] R¹ is selected from R, F, Cl, N(R⁸)₂, OR, NRCOR, NRCON(R⁸)₂,CON(R⁸)₂, SO₂R, NRSO₂R, or SO₂N(R⁸)₂;

[0067] T is selected from a valence bond or a linker group; each R isindependently selected from hydrogen or an optionally substitutedaliphatic group having one to six carbons;

[0068] R² is selected from hydrogen, CN, halogen, or an optionallysubstituted group selected from aryl, aralkyl, heteroaryl, heterocyclyl,acyclic aliphatic chain group having one to six carbons, or a cyclicaliphatic group having three to ten carbons;

[0069] R³ is selected from C₁-C₄ aliphatic, OH, O(C₁-C₄ aliphatic),N(C₁-C₄ aliphatic)₂, F, Cl, or CN;

[0070] Q is a valence bond, J, or an optionally substituted C₁₋₆alkylidene chain wherein up to two nonadjacent carbons of the alkylidenechain are each optionally and independently replaced by J;

[0071] J is selected from —C(═O)—, —CO₂—, —C(O)C(O)—, —NRCONR⁸—,—N(R)N(R⁸)—, —C(═O)NR⁸—, —NRC(═O)—, —O—, —S—, —SO—, 'SO₂—, —N(R)O—,—ON(R⁸)—, —OC(═O)N(R⁸)—, —N(R)COO—, —SO₂N(R⁸)—, —N(R)SO₂—, or —N(R⁸)—;

[0072] R⁴ is selected from —R⁸, —R⁵, —NH₂, —NHR⁵, —N(R⁵)₂, or—NR⁵(CH₂)_(y)N(R⁵)₂;

[0073] each R⁵ is independently selected from R⁶, R⁷, —(CH₂)_(y)CH(R⁶)(R⁷), —(CH₂)_(y)R⁶, —(CH₂)_(y)CH(R⁶)₂, —(CH₂)_(y)CH(R⁷)₂, or—(CH₂)_(y)R⁷;

[0074] y is 0-6;

[0075] each R⁶ is an optionally substituted group independently selectedfrom an aliphatic, aryl, aralkyl, aralkoxy, heteroaryl, heteroarylalkyl,heteroarylalkoxy, heterocyclyl, heterocyclylalkyl, orheterocyclylalkoxy, group;

[0076] each R⁷ is independently selected from an optionally substitutedhydroxyalkyl, alkoxyalkyl, aryloxyalkyl, or alkoxycarbonyl; and

[0077] each R⁸ is independently selected from R or two R⁸ on the samenitrogen taken together with the nitrogen optionally form a four toeight membered, saturated or unsaturated heterocyclic ring having one tothree heteroatoms.

[0078] Compounds of formula I illustrate selected examples of compoundsthat were designed to contain the pharmacophoric features of the presentinvention. The pyrazole ring of formula I satisfies the Grp2requirements as described above. R² is attached to Grp2 by attachmentmeans T and satisfies the Grp1 requirements; Ht is attached to Grp2 by avalence bond and satisfies the Grp3 requirements.

[0079] More specific examples of compounds having the desiredpharmacophore features are shown below for the Ht ring being pyrrol-3-yl(II-A), pyrazol-3-yl (II-B), [1,2,4]triazol-3-yl (II-C),[1,2,3]triazol-4-yl (II-D and II-E), and tetrazol-5-yl (II-F):

[0080] wherein R¹⁻⁴, T, and Q are as described above.

[0081] Preferred compounds of formulae II-A, II-B, II-C, II-D, II-E, andII-F include those having one or more, and most preferably all, of thefollowing features: (a) Q is —CO—, —CO₂—, or —CONH—; (b) T is a valencebond; (c) R¹ is hydrogen or NHR; (d) R² is an optionally substitutedaryl ring, more preferably an optionally substituted phenyl ring; (e) R³is hydrogen; (f) R⁴ is selected from R⁵, —NHR⁵, —N(R⁵)₂, —NR⁵R⁶,—NHCHR⁵R⁶, or —NHCH₂R⁵; and/or (g) R⁵ is an optionally substituted groupselected from aryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl,heterocyclylalkyl group, (CH₂)_(y)R⁶, ((CH₂)_(y)R⁷, or (CH₂)_(y)CH(R⁶)(R⁷).

[0082] Examples of substitutions of the R² phenyl group include halo,nitro, alkoxy, and amino.

[0083] When R⁴ is R⁵, examples of preferred R⁵ groups includepyrrolidin-1-yl, morpholin-1-yl, piperidin-1-yl, and piperazin-1-ylwherein each group is optionally substituted. When R⁴ is —NHR⁵ or—N(R⁵)₂, preferred R⁵ groups further include (CH₂)_(y)R⁶, (CH₂)_(y)R⁷,and (CH₂)_(y)CH (R⁶) (R⁷). Examples of preferred R⁶ and R⁷ includepyridin-3-yl, pyridin-4-yl, imidazolyl, furan-2-yl,tetrahydrofuran-2-yl, cyclohexyl, phenyl, —CH₂OH, —(CH₂)₂OH, isopropyl,wherein each group is optionally substituted.

[0084] Exemplary structures of formula II-A, wherein R¹ ans R³ are eachH, are set forth in Table 3 below. TABLE 3 Compounds II-A No. II-A- T—R²Q—R⁴ 1 phenyl CON(Me)₂ 2 phenyl CO₂Et 3 3-NO₂-phenyl CONHNH₂ 4 phenylCO(pyrrolidin-1-yl) 5 phenyl CONHCH₂(Ph) 6 3-NO₂-phenyl CO₂Et 74-Cl-phenyl CO₂Et 8 4-OMe-phenyl CO₂Et 9 3-NH₂-phenyl CO₂Et 103-OMe-phenyl CO₂Et 11 4-F-phenyl CO₂Et 12 4-NO₂-phenyl CO₂Et 133-Cl-phenyl CO₂Et 14 3-F-phenyl CO₂Et 15 phenyl CO₂H 16 4-NH₂-phenylCO₂Et 17 phenyl CONHCH₂CH₂N(Me)₂ 18 phenyl CONHCH₂(pyridin-3-yl) 19phenyl CO(morpholin-1-yl) 20 phenyl CONH(isopropyl) 21 phenylCO(4-Me-piperazin-1-yl) 22 phenyl CONHCH₂(furan-2-yl) 23 3-OMe-phenylCONMe₂ 24 3-OMe-phenyl CO(pyrrolidin-1-yl) 25 3-OMe-phenylCONHCH₂CH₂N(Me)₂ 26 3-OMe-phenyl CONHCH₂(pyridin-3-yl) 27 3-OMe-phenylCO(morpholin-1-y1) 28 3-OMe-phenyl CONH(isopropyl) 29 3-OMe-phenylCO(4-Me-piperazin-1-yl) 30 3-OMe-phenyl CONHCH₂(furan-2-yl) 314-NH₂-phenyl CO₂Et 32 H CONMe₂ 33 H CO(pyrrolidin-1-yl) 343-(AcNH)-phenyl CO₂Et 35 4-(AcNH)-phenyl CO₂Et 36 3-(AcNH)-phenyl CO₂Et37 4-(AcNH)-phenyl CO₂Et 38 3-Cl-phenyl CON(H)Bn 39 3,5-Cl₂-phenyl

CONH(2-OH-1-Ph-ethyl) 40 5-Br-phenyl CONH(3,4-F₂-phenyl) 41 5-Cl-phenylCONH(2-OH-1-Ph-ethyl) 42 4-OH,3-I,5-nitrophenyl CONH(2-OH-1-Ph-ethyl) 435-Br-phenyl

CONH(2,3-dihydro-benzofuran-5-yl) 44 3-NH₂,4-OH,5-I-phenylCONH(2-OH-1-Ph-ethyl) 45 5-Br-phenyl CONH(2-OH-1-Ph-ethyl) 465-Br-phenyl CONHCH₂(3-MeO-phenyl) 47 5-Br-phenyl CONHCH₂(3-CF₃-phenyl)48 3,5-Cl₂-phenyi CONHCH₂(pyrid-4-yl) 49 5-CF₃-phenylCONH(2-OH-1-Ph-ethyl) 50 5-Cl-phenyl CONHCH₂Ph 51 3,5-Cl₂-phenylCONHOCH₂Ph 52 4-OH,3-I,5-nitrophenyl CONHCH₂Ph 53 5-Cl-phenylCONHCH₂(pyrid-4-yl) 54 4,5-Cl₂-phenyl CONHOCH₂Ph 55 5-Br-phenylCONHCH₂(4-SO₂Me-phenyl) 56 5-Br-phenyl CONHNH(3-CF₃-phenyl) 575-Cl-phenyl CONHOCH₂Ph 58 5-Br-phenyl

CONHCH₂(5-Me-furan-2-yl) 59 5-Br-phenyl

CONH(3-OH-1-Ph-phenyl) 60 5-Br-phenyl CONHCH₂(2-Me-phenyl) 614,5-Cl₂-phenyl CONHCH₂(pyrid-4-yl) 62 5-Br-phenyl CONH(1-Ph-propyl) 635-F-phenyl CONHCH₂Ph 64 4,5-Cl₂-phenyl

CON(Me)(Et) 65 5-Br-phenyl

CONHCH₂(2-OH-cyclohexyl) 66 3,5-Cl₂-phenyl CON(Me)(Et) 67 5-Cl-phenylCONHCH₂(pyrid-3-yl) 68 5-Br-phenyl CONHCH₂(3,5-OMe₂-phenyl) 695-Br-phenyl CONNCH₂(2-OMe-phenyl) 70 4-F-5-Cl-phenyl CONHCH₂(pyrid-4-yl)71 4-F-5-Cl-phenyl CON(Me)(Et) 72 5-Br-phenyl CONH(2-OH-1-Ph-ethyl) 735-NH₂-phenyl CONHCH₂Ph 74 4,5-Cl₂-phenyl CONHCH₂(pyrid-3-yl) 755-Me-phenyl CONH(2-OH-1-Ph-ethyl) 76 3,5-Cl₂-phenyl CONHCH₂(pyrid-3-yl)77 4-F-5-Cl-phenyl CONHOCH₂Ph 78 3,5-Cl₂-phenylCONHCH₂(tetrahydrofuran-2-yl) 79 5-NO₂-phenyl CONHCH₂Ph 80 5-F-phenylCONHCH₂(pyrid-4-yl) 81 5-Cl-6-F-phenyl CON(Me)(Et) 82 2-F-3-Cl-phenylCONHOCH₂Ph 83 5-Br-phenyl

CON(Me)(CH₂Ph) 84 5-Cl-phenyl CONHCH₂(tetrahydrofuran-2-yl) 854,5-F₂-phenyl CONHOCH₂Ph 86 5-Br-phenyl CONH(3-OH-1-Ph-propyl) 875-Br-phenyl

88 4,5-F₂-phenyl CONHCH₂(pyrid-4-yl) 89 5-F-phenyl CONHOCH₂Ph 905-Me-phenyl CONHCH₂Ph 91 5-Br-phenyl

CONH(1-MeOH-2-Ph-ethyl) 92 4-Cl-phenyl CONHCH₂Ph 93 5-Cl-phenylCON(Me)(Et) 94 5-Br-phenyl CONHCH₂(4-SO₂NH₂-phenyl) 95 5-OH-phenylCONHCH₂Ph 96 5-Me-phenyl CONHCH₂(pyrid-4-yl) 97 Phenyl CONHCH₂Ph 982,5-F₂-phenyl CONHCH₂(pyrid-4-yl) 99 4-Cl-phenyl CONHOCH₂Ph 1004-F-5-Cl-phenyl CONHCH₂(tetrahydrofuran-2-yl) 101 4-F-5-Cl-phenylCONHCH₂(pyrid-3-yl) 102 5-Br-phenyl

CO(4-OH-4-Ph-piperidin-1-yl) 103 5,6-F₂-phenyl CONHOCH₂Ph 1045-Cl-phenyl CO(morpholin-1-yl) 105 5-Br-phenyl

106 2-F-3-Cl-phenyl CONHCH₂(tetrahydrofuran-2-yl) 107 4-F-5-Cl-phenylCO(morpholin-1-yl) 108 4-F-5-Cl-phenyl CON(Me)(Et) 109 5-Br-phenylCONHCH₂(4-NH₂-phenyl) 110 5-Br-phenyl

111 4-F-phenyl CONHCH₂Ph 112 3,5-Cl₂-phenyl CO(morphalin-1-y1) 1132,5-F₂-phenyl CONHOCH₂Ph 114 2-F-3-Cl-phenyl CONHCH₂(pyrid-3-yl) 1152-F-3-Cl-phenyl CONHCH₂(pyrid-4-yl) 116 4,5-F₂-phenylCONHCH₂(pyrid-3-yl) 117 4-OMe-phenyl CONHCH₂Ph 118 5-Br-phenylCONHCH₂(2,4,6-OMe₃-phenyl) 119 5-F-phenyl CONHCH₂(pyrid-3-yl) 1204,5-F₂-phenyl CONHCH₂(tetrahydrofuran-2-yl) 121 5-Cl-6-F-phenyl

122 5-Br-phenyl

123 5-Br-phenyl

124 5-Br-phenyl CONHCH₂(2,5-OMe₂-phenyl) 125 3,5-Cl₂-phenyl

226 5-Br-phenyl

127 4,5-Cl₂-phenyl CO(morpholin-1-yl) 128 5-Br-phenyl

129 2-F-3-Cl-phenyl CO(morpholin-1-yl) 130 5-Br-phenyl CONHCH₂CH₂OH 1315-NH₂-phenyl CONHCH₂Ph 132 5-MeOC(O)-phenyl CONHCH₂Ph 133 4-MeO-phenylCONHOCH₂Ph 134 phenyl CO(pyrrolidin-1-yl) 135 5-MeO-phenylCO(morpholin-1-y1) 136 5-Cl-phenyl CO(4-Me-piperidin-1-yl) 1375-NO₂-phenyl CONH₂NH₂ 138 5-Br-phenyl

139 5-Br-phenyl

140 5-Cl-phenyl CONHPh 141 5,6-F₂-phenyl CONHCH₂(pyrid-4-yl) 1425-Cl-phenyl

143 phenyl CON(Me)₂ 144 5-OMe-phenyl CO(pyrrolidin-1-yl) 1455-OMe-phenyl CONHCH₂(pyrid-3-yl) 146 4-F-phenyl CONHOCH₂Ph 1475-OMe-phenyl CONHCH₂(furan-2-yl) 148 5-NO₂-phenyl COOEt 149 phenylCONHCH₂(furan-2-yl) 150 phenyl CO(morpholin-1-yl) 151 5-Cl-phenyl COOEt152 5-Br-phenyl CONHMe 153 phenyl CONHCH₂(pyrid-3-yl) 154 5-OMe-phenylCON(Me)₂ 155 5-Cl-phenyl

CONH(2-OH-indan-1-yl) 156 5-Br-phenyl

157 5-Br-phenyl COOEt 158 phenyl CONH(iPr) 159 5-OMe-phenyl CONH(iPr)160 5-COOH-phenyl CONH(iPr) 161 5-Br-phenyl CONHO(iPr) 162 5-F-phenylCOOEt 163 5-OMe-phenyl CO(4-Me-piperidin-1-yl) 164 4-NH₂-phenyl COOEt165 4-NO₂-phenyl COOEt 166 pheny CO(4-Me-piperidin-1-yl) 167 4-Cl-phenylCOOEt 168 4-OMe-phenyl COOEt 169 pheny COOEt 170 5-OMe-phenyl COOEt 1714-F-phenyl COOEt 172 5-NH₂-phenyl COOEt 173 5-Cl-phenyl COOH 1745-Cl-phenyl

175 5-Cl-phenyl

176 5-OMe-phenyl CONHCH₂(pyrid-4-yl) 177 3,5-(OMe)₂-phenylCONHCH₂(pyrid-4-yl) 178 4-F-phenyl CONHCH₂(pyrid-3-yl) 179 4-OMe-phenylCONHCH₂(pyrid-3-yl) 180 2,5-(OMe)₂-phenyl CONHCH₂(pyrid-3-yl) 1812,5-F₂-phenyl CONHCH₂(pyrid-3-yl) 182 4-F-phenylCONHCH₂(tetrahydrofuran-2-yl) 183 4-OMe-phenylCONHCH₂(tetrahydrofuran-2-yl) 184 5-F-phenylCONHCH₂(tetrahydrofuran-2-yl) 185 5-OMe-phenylCONHCH₂(tetrahydrofuran-2-yl) 186 2,5-(OMe)₂-phenylCONHCH₂(tetrahydrofuran-2-yl) 187 5,6-F₂-phenylCONHCH₂(tetrahydrofuran-2-yl) 188 2,5-F₂-phenylCONHCH₂(tetrahydrofuran-2-yl) 189 4-F-phenylCONHCH₂(1-Et-pyrrolidin-2-yl) 190 4-OMe-phenylCONHCH₂(1-Et-pyrrolidin-2-yl) 191 5-F-phenylCONHCH₂(1-Et-pyrrolidin-2-yl) 192 5-OMe-phenylCONHCH₂(1-Et-pyrrolidin-2-yl) 193 3,6-(OMe)₂-phenylCONHCH₂(1-Et-pyrrolidin-2-yl) 194 4,5-F₂-phenylCONHCH₂(1-Et-pyrrolidin-2-yl) 195 5,6-F₂-phenylCONHCH₂(1-Et-pyrrolidin-2-yl) 196 3,6-F₂-phenylCONHCH₂(1-Et-pyrrolidin-2-yl) 197 4-F-phenyl CO(morpholin-1-yl) 1984-OMe-phenyl CO(morpholin-1-yl) 199 5-F-phenyl CO(morpholin-1-yl) 2002,5-(OMe)₂-phenyl CO(morpholin-1-yl) 201 4,5-F₂-phenylCO(morpholin-1-yl) 202 5,6-F₂-phenyl CO(morpholin-1-yl) 2032,5-F₂-phenyl CO(morpholin-1-yl) 204 4-F-phenyl CO(4-Me-piperidin-1-yl)205 4-OMe-phenyl CO(4-Me-piperidin-1-yl) 206 5-F-phenylCO(4-Me-piperidin-1-yl) 207 2,5-(OMe)₂-phenyl CO(4-Me-piperidin-1-yl)208 4,5-F₂-phenyl CO(4-Me-piperidin-1-yl) 209 5,6-F₂-phenylCO(4-Me-piperidin-1-yl) 210 3,6-F₂-phenyl CO(4-Me-piperidin-1-yl) 2114-Cl-phenyl CONHCH₂(pyrid-4-yl) 212 4,5-(OMe)₂-phenylCONHCH₂(pyrid-4-yl) 213 4-benzo[1,3]dioxo-5-yl CONHCH₂(pyrid-4-yl) 2144-Cl-phenyl CONHCH₂(pyrid-3-yl) 215 4,5-(OMe)₂-phenylCONHCH₂(pyrid-3-yl) 216 4-benzo[1,3]diaxo-5-yl CONHCH₂(pyrid-3-yl) 2174-Cl-phenyl CONHCH₂(tetrahydrofuran-2-yl) 218 4,5-(OMe)₂-phenylCONHCH₂(tetrahydrofuran-2-yl) 219 4-benzo[1,3]dioxo-5-ylCONHCH₂(tetrahydrofuran-2-yl) 220 4-Cl-phenylCONHCH₂(1-Et-pyrrolidin-2-yl) 221 4,5-Cl₂-phenylCONHCH₂(1-Et-pyrrolidin-2-yl) 222 5-Cl-6-F-phenylCONHCH₂(1-Et-pyrrolidin-2-yl) 223 4-F-5-Cl-phenylCONHCH₂(1-Et-pyrrolidin-2-yl) 224 4,5-(OMe)₂-phenylCONHCH₂(1-Et-pyrrolidin-2-yl) 225 4-benzo[1,3]dioxo-5-ylCONHCH₂(1-Et-pyrrolidin-2-yl) 226 3,5-Cl₂-phenylCONHCH₂(1-Et-pyrrolidin-2-yl) 227 4-Cl-phenyl CO(morpholin-1-yl) 2284,5-(OMe)₂-phenyl CO(morpholin-1-yl) 229 4-benzo[1,3]dioxo-5-ylCO(morpholin-1-yl) 230 4-Cl-phenyl CO(4-Me-piperidin-1-yl) 2314,5-Cl₂-phenyl CO(4-Me-piperidin-1-yl) 232 5-Cl-6-F-phenylCO(4-Me-piperidin-1-yl) 233 4-F-5-Cl-phenyl CO(4-Me-piperidin-1-yl) 2344,5-(OMe)₂-phenyl CO(4-Me-piperidin-1-yl) 235 4-benzo[1,3]dioxo-5-ylCO(4-Me-piperidin-1-yl) 236 3,5-Cl₂-phenyl CO(4-Me-piperidin-1-yl) 2375,6-F₂-phenyl CON(Me)(Et) 238 4-F-phenyl

239 5-OMe-phenyl

240 2,5-(OMe)₂-phenyl

241 4,5-F₂-phenyl

242 5,6-F₂-phenyl

243 3,6-F₂-phenyl

244 5-MeO-phenyl CONHOCH₂Ph 245 2,5-(OMe)₂-phenyl CONHOCH₂Ph 2465-F-phenyl

247 5-MeO-phenyl

248 4,5-F₂-phenyl

249 5,6-F₂-phenyl

250 5-Cl-phenyl

251 4-Cl-phenyl

252 4-Cl-phenyl

253 4,5-Cl₂-phenyl

254 4,5-Cl₂-phenyl

255 2-F-3-Cl-phenyl

256 4-F-5-Cl-phenyl

257 4-F-5-Cl-phenyl

258 4,5-(OMe)₂-phenyl CON(Me)(Et) 259 4,5-(OMe)₂-phenyl

260 4,5-(OMe)₂-phenyl CONHOCH₂Ph 261 4,5-(OMe)₂-phenyl

262 4-benzo[1,3]dioxo-5-yl CON(Me)(Et) 263 4-benzo[1,3]dioxo-5-yl

264 4-benzo[1,3]dioxo-5-yl CONHOCH₂Ph 265 4-benzo[1,3]dioxo-5-yl

266 3,5-Cl₂-phenyl

267 5-Br-phenyl

268 5-Br-phenyl

269 5-Br-phenyl

270 5-Br-phenyl

271 5-Br-phenyl

272 5-Br-phenyl

273 5-Br-phenyl

274 5-Br-phenyl

275 5-Br-phenyl

276 5-Br-phenyl

277 5-Br-phenyl

278 5-Br-phenyl

279 5-Br-phenyl

280 5-Br-phenyl CONH(CH₂)₂COOH 281 5-Br-phenyl

282 5-Br-phenyl CONHCH₂(4-COOH-phenyl) 283 5-Br-phenyl

284 5-Br-phenyl

285 3-NO₂-phenyl CONHCH₂phenyl 286 5-Cl-phenylCONHCH₂(1-Et-pyrrolidin-2-yl) 287 5-(N-Et-NHCO)-phenyl CONHCH₂phenyl 2885-Br-phenyl

289 5-NO₂-phenyl CONHCH₂(pyrid-4-yl) 290 5-Br-phenyl

291 5-F-phenyl CON(Me)(Et) 292 5-MeO-phenyl CON(Me)(Et) 293 5-Br-phenyl

294 5-Br-phenyl

295 5-Br-phenyl

296 5-Br-phenyl

297 phenyl CONH(CH₂)₂NMe₂ 298 5-MeO-phenyl CONH(CH₂)₂NMe₂ 2995-Br-phenyl CONHCH₂phenyl 300 3-Cl-phenyl

301 3-Cl-phenyl

302 3-Cl-phenyl

303 3-Cl-phenyl

304 3-Cl-phenyl

305 3-Cl-phenyl

306 3-Cl-phenyl

307 3-Cl-phenyl

308 3-Cl-phenyl

309 3-Cl-phenyl

310 3,5-Cl₂-phenyl

311 3-Br-5-CF₃-phenyl

312 3-Cl-phenyl

313 3,5-Cl₂-phenyl

314 3-Cl-4-CN-phenyl

315 3-Cl-4-CH₂OH-phenyl

316 3-Cl-4-CH₂NH₂-phenyl

317

318

319

320

321

322

323 CH₂Ph CON(Me)₂ 324 cyclopentylmethyl CO₂NHCH₂Ph 325 isopropyl CN 3263-Cl-phenyl NHCOCH₂Ph 327 3-Cl-phenyl NHSO₂-morpholin-1-yl 3283-Cl-phenyl NHCONHCH₂Ph 329 3-Cl-phenyl NHCO₂-tetrahydrofuran-2-yl 330CH₂Ph CONHCH₂Ph 331 Me CONHCH₂Ph 332 isopropyl CONHCH₂Ph 333 H CON(Me)₂

[0085] Exemplary structures of formulae II-B, II-C, II-D, and II-F,wherein R¹ and R³ are each H and T is a valence bond, are set forth inTable 4 below. TABLE 4 Compounds II-B, II-C, II-D, and II-F No.Structure II-B-1

II-B-2

II-B-3

IIB-4

II-B-5

II-B-6

II-B-7

II-B-8

II-C-1

II-C-2

II-C-3

II-D-1

II-D-2

II-D-3

II-D-4

II-F-1

II-F-2

II-F-3

[0086] It will be apparent to one skilled in the art that certaincompounds of this invention may exist in tautomeric forms, all suchtautomeric forms of the compounds being within the scope of theinvention. Examples of tautomeric forms of this invention include, butare not limited to, the tautomers shown below.

[0087] Unless otherwise stated, structures depicted herein are alsomeant to include all stereochemical forms of the structure; i.e., the Rand S configurations for each asymmetric center. Therefore, singlestereochemical isomers as well as enantiomeric and diastereomericmixtures of the present compounds are within the scope of the invention.Unless otherwise stated, structures depicted herein are also meant toinclude compounds which differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructures except for the replacement of a hydrogen by a deuterium ortritium, or the replacement of a carbon by a ¹³C— or ¹⁴C-enriched carbonare within the scope of this invention.

[0088] Other examples of compounds having the desired pharmacophorefeatures are shown below for the Ht ring being pyrrol-3-yl (III-A),pyrazol-3-yl (III-B), [1,2,4]triazol-3-yl (III-C), [1,2,3]triazol-4-yl(III-D and III-E), and tetrazol-5-yl (III-F):

[0089] wherein T, R, R², and R⁴ are as described above.

[0090] Preferred compounds of formulae III-A, III-B, Ill-C, III-D,III-E, and III-F include those having one or more, and most preferablyall, of the following features: (a) Q is —CO—, —CO₂—, or —CONH—; (b) Tis a valence bond; (c) R² is an optionally substituted aryl ring, morepreferably an optionally substituted phenyl ring; (d) R³ is hydrogen;(e) R⁴ is selected from R⁵, —NHR⁵, —N(R⁵)₂, —NR⁵R⁶, —NHCHR⁵R⁶, or 13NHCH₂R⁵; and/or (f) R⁵ is an optionally substituted group selected fromaryl, aralkyl, heteroaryl, heteroarylalkyl, heterocyclyl,heterocyclylalkyl group, (CH₂)_(y)R⁶, (CH₂)_(y)R⁷, or (CH₂)_(y)CH(R⁶)(R⁷).

[0091] Exemplary compounds of formula III-A, wherein R³ is H and T is avalence bond, are set forth in Table 5 below. TABLE 5 Compounds III-ANo. III-A- R R² Q—R⁴ 1 H phenyl CON(Me)₂ 2 H phenyl CO₂Et 3 H3-NO₂-phenyl CONHNH₂ 4 H phenyl CO(pyrrolidin-1-yl) 5 Me phenylCONHCH₂(Ph) 6 H 3-NO₂-phenyl CO₂Et 7 H 4-Cl-phenyl CO₂Et 8 Me4-OMe-phenyl CO₂Et 9 H 3-NH₂-phenyl CO₂Et 10 H 3-OMe-phenyl CO₂Et 11 H4-F-phenyl CO₂Et 12 H 4-NO₂-phenyl CO₂Et 13 Et 3-Cl-phenyl CO₂Et 14 H3-F-phenyl CO₂Et 15 H phenyl CO₂H 16 Me 3-Cl-phenylCONHCH₂(pyridin-4-yl) 17 H 5-Cl-phenyl

18 H 5-F-phenyl CONHCH₂(tetrahydrofuran-2-yl) 19 Me 5,6-F₂-phenylCO(4-Me-piperidin-1-yl) 20 H 4-Cl-phenyl CONHCH₂(pyrid-4-yl) 21 H4,5-(OMe)₂-phenyl

22 Me 4,5-Cl₂-phenyl

23 H 3-Cl-phenyl

24 H 3-Cl-phenyl

25 Me 3,5-Cl₂-phenyl

26 H

[0092] Exemplary compounds of formula III-B, III-C, III-D, and III-F,wherein R³ is H and T is a valence bond, are set forth in Table 6 below.TABLE 6 Compounds III-B, III-C, III-D, and III-F No. Structure III-B-1

III-B-2

III-B-3

III-B-4

III-B-5

III-C-1

III-C-2

III-C-3

III-D-1

III-D-2

[0093]

[0094] Reagents and conditions: (a) PhCH₂COCl, AlCl₃, CH₂Cl₂, 2 hours,RT (b) DMF, 24 hrs, room temperature (c) (Me₂N)₂-Ot-Bu, THF, 24 hrs,room temperature (d) H₂NNH₂, EtOH, 12 hours, reflux

[0095] Scheme I above shows the general synthetic method that was usedfor preparing compounds of formula II-A of this invention. In step (a),the optionally substituted benzoyl chloride was combined with compound 1in dichloromethane and aluminum trichloride to form compound 2. A widevariety of substitutions on the phenyl ring are amenable to thisreaction. Examples of suitable R² groups include, but are not limitedto, those set forth in Table 3 above.

[0096] The formation of amide 4 was achieved by treating compound 2 witheach of a variety of amines 3 in DMF. When amine 3 was a primary amine,the reaction proceeded at ambient temperature. When amine 3 was asecondary amine, the reaction was heated at 50° C. to achieve completereaction and afford amide 4.

[0097] The formation of enamine 5 at step (c) was achieved by treatingamide 4 with (Me₂N)₂-Ot-Bu at ambient temperature. Alternatively, thereaction to form enamine 5 at step (c) was also achieved by usingdimethylformamide-dimethylacetal (DMF-DMA). The reaction using DMF-DMArequires elevated temperature to afford enamine 5 whereas using(Me₂N)₂-OtBu has the advantage of proceeding at ambient temperature toafford the enamine 5 in higher purity.

[0098] The formation of the pyrazole compound 6 at step (d) was achievedby the treatment of enamine 5 with hydrazine hydrate at elevatedtemperature. The compounds of formula II-A synthesized by this method,as exemplified in Table 3, were isolated by preparatory HPLC (reversephase, 10→90% MeCN in water over 15 minutes). The details of theconditions used for producing these compounds are set forth in theExamples.

[0099] Compounds of formula II-B may be prepared according to a modifiedmethod of Finar, I. L., J. Chem. Soc., (1955), pp. 1205, as shown inScheme II below for the preparation of compound II-B-6.

[0100] Compounds of formula II-C may be prepared according to themethods of Clitherow, J. W., et al, Bioorg. Med. Chem. Lett., (1996) pp.833-838, as shown in Scheme III below for the preparation of compoundII-C-3.

[0101] Compounds of formula II-D may be prepared according to themethods of Beck, G., et al, Chem. Ber., (1973) pp. 106, as shown inScheme IV below for the preparation of compound II-D-4.

[0102] Compounds of formula II-F may be prepared according to themethods of Kaltenbronn, J. S., et al, Eur. J. med. Chem., (1997) pp.425-431, and Norman, M. H., et al, (1995) pp. 4670-4678, as shown inScheme V below for the preparation of compound II-F-3.

[0103] Compounds of formula III-A may be prepared in general accordingto the methods of Jira, T., et al, Pharmazie, pp. 401-406 (1994) asshown in Scheme VI below for the synthesis of compound III-A-16.

[0104] Reagents and conditions: (a) 3-Cl-PhCH₂COCl, AlCl₃, CH₂Cl₂, 2hours, RT (b) DMF, 24 hrs, room temperature (c) NBS, CCl₄, reflux (d)iPrOH, reflux (e) formic acid, reflux, 2 hours.

[0105] Using compound III-A-16 as an example, Scheme VI above shows ageneral synthetic method that may be used for preparing compounds offormula III-A. This method is modified from that of Jira, T., et al,Pharmazie, pp. 401-406 (1994). Compounds of formula III-A may also beprepared by methods similar to those of Woller, J., et al, Pharmazie,pp. 937-940 (1996), Rychmans, T., et al, Tetrahedron, pp. 1729-1734(1997), and Tupper, D. E., et al, Synthesis, pp. 337-341 (1997).

[0106] According to another embodiment, the invention provides a methodof inhibiting kinase activity in a biological sample. This methodcomprises the step of contacting said biological sample with a compoundof this invention.

[0107] The term “biological sample”, as used herein includes cellcultures or extracts thereof; biopsied material obtained from a mammalor extracts thereof; and blood, saliva, urine, feces, semen, tears, orother body fluids or extracts thereof. The term “biological sample” alsoincludes living organisms, in which case “contacting a compound of thisinvention with a biological sample” is synonymous with the term“administrating said compound (or composition comprising said compound)to a mammal.”

[0108] One aspect of this invention relates to a method for treating adisease state in mammals that is alleviated by treatment with a proteinkinase inhibitor, especially an ERK inhibitor, which method comprisesadministering to a mammal in need of such a treatment a therapeuticallyeffective amount of a compound having the pharmacophoric features ofthis invention.

[0109] The method is particularly useful for treating a disease statethat is alleviated by the use of an inhibitor of one or more enzymesselected from ERK or JAK.

[0110] One embodiment of this method comprises administering a compoundof formula I, preferably a compound of formula II-A. Another embodimentcomprises administering a compound of formula III, preferably a compoundof formula III-A, and most preferably, a compound listed in Tables 3-6.Pharmaceutical compositions useful for such methods are described below.

[0111] The activity of the compounds as protein kinase inhibitors, forexample as ERK inhibitors, may be assayed in vitro, in vivo or in a cellline. In vitro assays include assays that determine inhibition of eitherthe kinase activity or ATPase activity of activated ERK. Alternate invitro assays quantitate the ability of the inhibitor to bind to ERK andmay be measured either by radiolabelling the inhibitor prior to binding,isolating the inhibitor/ERK complex and determining the amount ofradiolabel bound, or by running a competition experiment where newinhibitors are incubated with ERK bound to known radioligands. One mayuse any type or isoform of ERK, depending upon which ERK type or isoformis to be inhibited.

[0112] The protein kinase inhibitors, particularly ERK inhibitors, orpharmaceutical salts thereof may be formulated into pharmaceuticalcompositions for administration to animals or humans. Thesepharmaceutical compositions, which comprise an amount of the proteininhibitor effective to treat or prevent an ERK-mediated condition and apharmaceutically acceptable carrier, are another embodiment of thepresent invention.

[0113] The term “ERK-mediated condition”, as used herein means anydisease or other deleterious condition in which ERK is known to play arole. Such conditions include, without limitation, cancer, stroke,diabetes, hepatomegaly, cardiovascular disease including cardiomegaly,Alzheimer's disease, cystic fibrosis, viral disease, autoimmunediseases, atherosclerosis, restenosis, psoriasis, allergic reactionsincluding asthma, inflammation, neurological disorders andhormone-related diseases. The term “cancer” includes, but is not limitedto the following cancers: breast; ovary; cervix; prostate; testis,genitourinary tract; esophagus; larynx, glioblastoma; neuroblastoma;stomach; skin, keratoacanthoma; lung, epidermoid carcinoma, large cellcarcinoma, small cell carcinoma, lung adenocarcinoma; bone; colon,adenoma; pancreas, adenocarcinoma; thyroid, follicular carcinoma,undifferentiated carcinoma, papillary carcinoma; seminoma; melanoma;sarcoma; bladder carcinoma; liver carcinoma and biliary passages; kidneycarcinoma; myeloid disorders; lymphoid disorders, Hodgkin's, hairycells; buccal cavity and pharynx (oral), lip, tongue, mouth, pharynx;small intestine; colon-rectum, large intestine, rectum; brain andcentral nervous system; and leukemia.

[0114] In addition to the compounds of this invention, pharmaceuticallyacceptable derivatives or prodrugs of the compounds of this inventionmay also be employed in compositions to treat or prevent theabove-identified disorders.

[0115] A “pharmaceutically acceptable derivative or prodrug” means anypharmaceutically acceptable salt, ester, salt of an ester or otherderivative of a compound of this invention which, upon administration toa recipient, is capable of providing, either directly or indirectly, acompound of this invention or an inhibitorily active metabolite orresidue thereof. Particularly favored derivatives or prodrugs are thosethat increase the bioavailability of the compounds of this inventionwhen such compounds are administered to a mammal (e.g., by allowing anorally administered compound to be more readily absorbed into the blood)or which enhance delivery of the parent compound to a biologicalcompartment (e.g., the brain or lymphatic system) relative to the parentspecies.

[0116] Pharmaceutically acceptable prodrugs of the compounds of thisinvention include, without limitation, esters, amino acid esters,phosphate esters, metal salts and sulfonate esters.

[0117] Pharmaceutically acceptable salts of the compounds of thisinvention include those derived from pharmaceutically acceptableinorganic and organic acids and bases. Examples of suitable acid saltsinclude acetate, adipate, alginate, aspartate, benzoate,benzenesulfonate, bisulfate, butyrate, citrate, camphorate,camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate,ethanesulfonate, formate, fumarate, glucoheptanoate, glycerophosphate,glycolate, hemisulfate, heptanoate, hexanoate, hydrochloride,hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate,oxalate, palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate,picrate, pivalate, propionate, salicylate, succinate, sulfate, tartrate,thiocyanate, tosylate and undecanoate. Other acids, such as oxalic,while not in themselves pharmaceutically acceptable, may be employed inthe preparation of salts useful as intermediates in obtaining thecompounds of the invention and their pharmaceutically acceptable acidaddition salts.

[0118] Salts derived from appropriate bases include alkali metal (e.g.,sodium and potassium), alkaline earth metal (e.g., magnesium), ammoniumand N⁺(C₁₋₄ alkyl)₄ salts. This invention also envisions thequaternization of any basic nitrogen-containing groups of the compoundsdisclosed herein. Water or oil-soluble or dispersible products may beobtained by such quaternization.

[0119] Pharmaceutically acceptable carriers that may be used in thesepharmaceutical compositions include, but are not limited to, ionexchangers, alumina, aluminum stearate, lecithin, serum proteins, suchas human serum albumin, buffer substances such as phosphates, glycine,sorbic acid, potassium sorbate, partial glyceride mixtures of saturatedvegetable fatty acids, water, salts or electrolytes, such as protaminesulfate, disodium hydrogen phosphate, potassium hydrogen phosphate,sodium chloride, zinc salts, colloidal silica, magnesium trisilicate,polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol,sodium carboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat.

[0120] The compositions of the present invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes subcutaneous, intravenous, intramuscular,intra-articular, intra-synovial, intrasternal, intrathecal,intrahepatic, intralesional and intracranial injection or infusiontechniques. Preferably, the compositions are administered orally,intraperitoneally or intravenously.

[0121] Sterile injectable forms of the compositions of this inventionmay be aqueous or oleaginous suspension. These suspensions may beformulated according to techniques known in the art using suitabledispersing or wetting agents and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally-acceptable diluent or solvent,for example as a solution in 1,3-butanediol. Among the acceptablevehicles and solvents that may be employed are water, Ringer's solutionand isotonic sodium chloride solution. In addition, sterile, fixed oilsare conventionally employed as a solvent or suspending medium. For thispurpose, any bland fixed oil may be employed including synthetic mono-or di-glycerides. Fatty acids, such as oleic acid and its glyceridederivatives are useful in the preparation of injectables, as are naturalpharmaceutically-acceptable oils, such as olive oil or castor oil,especially in their polyoxyethylated versions. These oil solutions orsuspensions may also contain a long-chain alcohol diluent or dispersant,such as carboxymethyl cellulose or similar dispersing agents which arecommonly used in the formulation of pharmaceutically acceptable dosageforms including emulsions and suspensions. Other commonly usedsurfactants, such as Tweens, Spans and other emulsifying agents orbioavailability enhancers which are commonly used in the manufacture ofpharmaceutically acceptable solid, liquid, or other dosage forms mayalso be used for the purposes of formulation.

[0122] The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In thecase of tablets for oral use, carriers commonly used include lactose andcorn starch. Lubricating agents, such as magnesium stearate, are alsotypically added. For oral administration in a capsule form, usefuldiluents include lactose and dried cornstarch. When aqueous suspensionsare required for oral use, the active ingredient is combined withemulsifying and suspending agents. If desired, certain sweetening,flavoring or coloring agents may also be added.

[0123] Alternatively, the pharmaceutical compositions of this inventionmay be administered in the form of suppositories for rectaladministration. These can be prepared by mixing the agent with asuitable non-irritating excipient which is solid at room temperature butliquid at rectal temperature and therefore will melt in the rectum torelease the drug. Such materials include cocoa butter, beeswax andpolyethylene glycols.

[0124] The pharmaceutical compositions of this invention may also beadministered topically, especially when the target of treatment includesareas or organs readily accessible by topical application, includingdiseases of the eye, the skin, or the lower intestinal tract. Suitabletopical formulations are readily prepared for each of these areas ororgans.

[0125] Topical application for the lower intestinal tract can beeffected in a rectal suppository formulation (see above) or in asuitable enema formulation. Topically-transdermal patches may also beused.

[0126] For topical applications, the pharmaceutical compositions may beformulated in a suitable ointment containing the active componentsuspended or dissolved in one or more carriers. Carriers for topicaladministration of the compounds of this invention include, but are notlimited to, mineral oil, liquid petrolatum, white petrolatum, propyleneglycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax andwater. Alternatively, the pharmaceutical compositions can be formulatedin a suitable lotion or cream containing the active components suspendedor dissolved in one or more pharmaceutically acceptable carriers.Suitable carriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol,2-octyldodecanol, benzyl alcohol and water.

[0127] For ophthalmic use, the pharmaceutical compositions may beformulated as micronized suspensions in isotonic, pH adjusted sterilesaline, or, preferably, as solutions in isotonic, pH adjusted sterilesaline, either with or without a preservative such as benzylalkoniumchloride. Alternatively, for ophthalmic uses, the pharmaceuticalcompositions may be formulated in an ointment such as petrolatum.

[0128] The pharmaceutical compositions of this invention may also beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents.

[0129] The amount of ERK inhibitor that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated, the particular mode of administration. Preferably, thecompositions should be formulated so that a dosage of between 0.01-100mg/kg body weight/day of the inhibitor can be administered to a patientreceiving these compositions.

[0130] It should also be understood that a specific dosage and treatmentregimen for any particular patient will depend upon a variety offactors, including the activity of the specific compound employed, theage, body weight, general health, sex, diet, time of administration,rate of excretion, drug combination, and the judgment of the treatingphysician and the severity of the particular disease being treated. Theamount of inhibitor will also depend upon the particular compound in thecomposition.

[0131] The protein kinase inhibitors of this invention, orpharmaceutical salts thereof, may be formulated into pharmaceuticalcompositions for administration to animals, especially mammals orhumans. These pharmaceutical compositions effective to treat or preventa protein kinase-mediated condition which comprise the protein kinaseinhibitor in an amount sufficient to detectably inhibit protein kinaseactivity and a pharmaceutically acceptable carrier, are anotherembodiment of the present invention. The term “detectably inhibit”, asused herein means a measurable change in activity between a samplecontaining said inhibitor and a sample containing only a protein kinase.

[0132] According to another embodiment, the invention provides methodsfor treating or preventing a ERK-mediated condition comprising the stepof administering to a patient one of the above-described pharmaceuticalcompositions. The term “patient”, as used herein, means an animal,preferably a mammal, and most preferably a human.

[0133] Preferably, that method is used to treat or prevent a conditionselected from cancers such as cancers of the breast, colon, prostate,skin, pancreas, brain, genitourinary tract, lymphatic system, stomach,larynx and lung, including lung adenocarcinoma and small cell lungcancer, stroke, diabetes, hepatomegaly, cardiomegaly, Alzheimer'sdisease, cystic fibrosis, and viral disease, or any specific disease ordisorder described above.

[0134] The kinase inhibitors of this invention or pharmaceuticalcompositions thereof may also be incorporated into compositions forcoating an implantable medical device, such as prostheses, artificialvalves, vascular grafts, stents and catheters. Vascular stents, forexample, have been used to overcome restenosis (re-narrowing of thevessel wall after injury). However, patients using stents or otherimplantable devices risk clot formation or platelet activation. Theseunwanted effects may be prevented or mitigated by pre-coating the devicewith a composition comprising a kinase inhibitor. Compositionscomprising a kinase inhibitor of this invention and a suitable carrieror coating are another embodiment of the present invention.

[0135] Suitable coatings and the general preparation of coatedimplantable devices are described in U.S. Pat. Nos. 6,099,562;5,886,026; and 5,304,121. The coatings are typically biocompatiblepolymeric materials such as a hydrogel polymer, polymethyldisiloxane,polycaprolactone, polyethylene glycol, polylactic acid, ethylene vinylacetate, and mixtures thereof. The coatings may optionally be furthercovered by a suitable topcoat of fluorosilicone, polysaccarides,polyethylene glycol, phospholipids or combinations thereof to impartcontrolled release characteristics in the composition. Implantabledevices coated with a kinase inhibitor of this invention are anotherembodiment of the present invention.

[0136] Depending upon the particular condition, or disease state, to betreated or prevented, additional therapeutic agents, which are normallyadministered to treat or prevent that condition, may be administeredtogether with the inhibitors of this invention. For example,chemotherapeutic agents or other anti-proliferative agents may becombined with the inhibitors of this invention to treat proliferativediseases and cancer. Examples of known chemotherapeutic agents include,but are not limited to, adriamycin, dexamethasone, vincristine,cyclophosphamide, fluorouracil, topotecan, taxol, interferons, andplatinum derivatives.

[0137] Other examples of agents the inhibitors of this invention mayalso be combined with include, without limitation, anti-inflammatoryagents such as corticosteroids, TNF blockers, IL-1 RA, azathioprine,cyclophosphamide, and sulfasalazine; immunomodulatory andimmunosuppressive agents such as cyclosporin, tacrolimus, rapamycin,mycophenolate mofetil, interferons, corticosteroids, cyclophophamide,azathioprine, and sulfasalazine; neurotrophic factors such asacetylcholinesterase inhibitors, MAO inhibitors, interferons,anti-convulsants, ion channel blockers, riluzole, and anti-Parkinsonianagents; agents for treating cardiovascular disease such asbeta-blockers, ACE inhibitors, diuretics, nitrates, calcium channelblockers, and statins; agents for treating liver disease such ascorticosteroids, cholestyramine, interferons, and anti-viral agents;agents for treating blood disorders such as corticosteroids,anti-leukemic agents, and growth factors; agents for treating diabetessuch as insulin, insulin analogues, alpha glucosidase inhibitors,biguanides, and insulin sensitizers; and agents for treatingimmunodeficiency disorders such as gamma globulin.

[0138] These additional agents may be administered separately, as partof a multiple dosage regimen, from the inhibitor-containing composition.Alternatively, these agents may be part of a single dosage form, mixedtogether with the inhibitor in a single composition.

[0139] In order that the invention described herein may be more fullyunderstood, the following examples are set forth. It should beunderstood that these examples are for illustrative purposes only andare not to be construed as limiting this invention in any manner.

EXAMPLES Example 1

[0140]

[0141] 2,2,2-Trichloro-1-(4-phenylacetyl-1H-pyrrol-2-yl)-ethanone (1):In a dry flask, phenylacetyl chloride (1 equivalent) was combined with2-trichloroacetyl pyrrole (1 equivalent) in a minimum amount ofdichloromethane (DCM). To the resulting solution, at ambienttemperature, was added aluminum trichloride (1 equivalent). After 2hours, the reaction mixture was applied directly onto a silica gelcolumn. Gradient elution with 10% ethyl acetate to 50% ethyl acetate inhexanes provided compound 1 in 60% yield. ¹H NMR (CDCl₃) □4.0 (s, 2H),7.1-7.35 (m, 7H), 9.7 (br s, NH). HPLC using method B (as describedbelow for Example 5) provided a retention time of 4.9 minutes. LC/MS(M+1) 330.2, (M−1) 328.1.

Example 2

[0142]

[0143] 4-Phenylacetyl-1H-pyrrole-2-carboxylic acid benzylamide (2): To asolution of compound 1 (1 equivalent) in DMF, at ambient temperature,was added benzylamine (1.2 equivalents). After 24 hours, the solvent wasevaporated and the crude product 2 was utilized without purification.HPLC using method B (as described below for Example 5) provided aretention time of 3.8 minutes. FIA/MS (M+1) 319.3, (M−1) 317.2.

Example 3

[0144]

[0145] 4-(3-Dimethylamino-2-phenyl-acryloyl)-1H-pyrrole-2-carboxylicacid benzylamide (3): To a solution of compound 2 (1 equivalent) in THF,at ambient temperature, was added (Me₂N)₂CHOt-Bu (3 equivalents). After24 hours, the solvent was evaporated and the crude product 3 wasutilized without purification. ¹H NMR (CDCl₃) □4.4 (s, 2H), 4.8 (s, NH),6.8-7.4 (m, 13H).

Example 4

[0146]

[0147] 4-(4-phenyl-1H-pyrazole-3-yl)-1H-pyrrole-2-carboxylic acidbenzylamide (II-5): To a solution of compound 3 (1 equivalent) inethanol, at ambient temperature, was added hydrazine hydrate (3equivalents) and the resulting mixture heated at reflux. After 12 hours,the solvent was evaporated and the crude product purified by preparatoryHPLC (reverse phase; 10→90% MeCN in water; 15 minutes) to afford thedesired compound II-5. LC/MS (M+1) 343.3, (M−1) 341.2.

Example 5

[0148] We have prepared other compounds of formula II by methodssubstantially similar to those described in the above Examples 1-4 andas illustrated in Scheme I. The characterization data for thesecompounds is summarized in Table 7 below and includes LC/MS, HPLC, and¹H NMR data.

[0149] For compounds where the HPLC Method is designated as “A”, thefollowing method was utilized: a gradient of water:MeCN, 0.1% TFA(95:5→0:100) was run over 22 minutes at 1 mL/min and 214 nm. Forcompounds where the HPLC Method is designated as “B”, the followingmethod was utilized: a gradient of water:MeCN, 0.1% TFA (90:10→0:100)was run over 8 minutes at 1 mL/min and 214 nm. Each of methods A and Butilize the YMC ODS-AQ 55 120A column with a size of 3.0×150 mm. Theterm “T_(ret)(min)” refers to the retention time, in minutes, associatedwith the compound using the designated HPLC method.

[0150] Where applicable, ¹H NMR data is also summarized in Table 7 belowwherein “Y” designates ¹H NMR data is available and was found to beconsistant with structure. Compound numbers correspond to the compoundnumbers listed in Table 3. TABLE 7 Characterization Data for SelectedCompounds Compound No II-A- M +1 M −1 HPLC Method T_(ret) (min) ¹H NMR41 407.4 405.4 A 8.6 Y 42 560.2 558.1 A 9.5 — 43 — — A 10.5 — 44 530.3528.2 A 6.3 — 45 — — A 9.8 — 46 — — A 10.6 — 50 377.4 — A 10.1 Y 52530.2 528.2 A 10.3 — 53 378.4 376.3 A 7.4 Y 56 490.2 488.1 A 10.8 — 58 —— A 10.46 — 59 — — A 9.1 — 63 361.4 359.3 A 9.5 Y 65 — — A 10.0 — 67378.4 376.3 A 7.4 Y 72 451.5 449.1 A 10.15 Y 80 374.4 372.3 A 6.6 — 83435.3 433.4 A 10.3 — 85 — — A 10.6 — 86 — — A 9.3 — 88 380.4 378.3 A 6.9— 89 — — A 10.5 — 91 — — A 9.6 — 92 377.4 375.3 A 10.2 Y 94 — — A 9.0 —97 342.1 — B 3.8 Y 98 380.4 378.3 A 6.7 — 102 — — A 10.3 — 103 — — A10.6 — 105 — — A 9.3 — 109 — — A 7.9 — 110 — — A 10.3 — 111 361.4 359.3A 9.4 Y 113 — — A 10.6 — 116 380.2 378.4 A 6.9 — 117 373.4 — A 9.0 Y 119362.4 371.4 A 6.5 — 120 373.4 371.4 A 8.2 — 122 — — A 10.8 — 123 — — A11.4 — 126 — — A 10.2 — 128 — — A 10.9 — 130 — — A 7.4 — 133 — — A 9.5 —134 306.1 — B 3.5 Y 135 353.4 351.4 A 7.7 — 137 313.3 311.2 A 6.4 Y 141380.4 378.3 A 6.7 — 143 280.1 — B 3.3 Y 144 336.4 — B 3.5 — 145 373.4 —B 2.8 — 146 — — A 10.5 — 147 362.4 — B 3.5 — 148 327.3 325.2 A 9.2 Y 149332.4 — B 3.5 — 150 322.4 — B 3.2 — 151 316.2 314.2 A 10.3 Y 152 — — A6.6 — 153 323.4 — B 2.3 — 154 343.4 — B 2.8 — 158 294.3 — B 3.4 — 159335.4 — B 2.7 — 161 389.3 387.2 A 8.9 — 162 300.3 298.2 A 9.5 Y 163366.5 364.4 B 6.0 — 164 297.3 — A 5.1 Y 165 322.3 325.2 A 9.7 Y 167316.2 314.2 A 10.0 Y 168 312.3 310.2 A 8.6 Y 169 281.1 — B 3.9 Y 170312.3 310.2 A 9.1 Y 171 300.3 298.2 A 9.4 Y 172 297.3 295.7 A 5.5 Y 174449.3 447.2 A 12.5 Y 175 477.3 475.3 A 14.0 Y 176 374.4 372.4 A 6.3 —178 362.4 360.0 A 6.6 — 179 374.4 372.4 A 6.3 — 180 404.4 402.4 A 6.4 —181 380.2 378.3 A 6.7 — 182 355.4 353.4 A 7.7 — 183 367.4 365.4 A 7.4 —184 355.4 353.4 A 7.9 — 185 367.4 365.3 A 7.5 — 186 397.4 395.4 A 7.1 —187 373.4 371.4 A 8.0 — 188 373.4 371.4 A 7.9 — 189 382.4 380.4 A 6.9 —190 394.4 392.4 A 6.7 — 191 382.4 380.4 A 7.0 — 192 394.5 392.4 A 6.7 —193 424.4 422.4 A 6.4 — 194 400.4 398.4 A 7.3 — 195 400.4 398.4 A 7.1 —196 400.4 398.4 A 7.2 — 197 341.3 339.2 A 7.5 — 198 353.4 351.4 A 7.1 —199 341.3 339.2 A 7.6 — 200 383.4 381.4 A 6.9 — 201 359.4 357.4 A 8.0 —202 359.4 357.4 A 7.8 — 203 359.4 357.4 A 7.7 — 204 354.4 352.4 A 6.2 —205 366.4 364.4 A 5.9 — 206 354.4 .52.4 A 5.6 — 207 396.4 394.4 A 5.9 —208 372.4 370.4 A 6.7 — 209 372.4 370.4 A 6.5 — 210 372.4 370.4 A 6.4 —237 — — A 9.8 — 238 — — A 11.6 — 239 — — A 11.3 — 240 — — A 7.5 — 241 —— A 12.0 — 242 — — A 11.7 — 243 — — A 11.6 — 244 389.4 387.3 A 10.2 —245 — — A 10.6 — 246 365.4 363.4 A 7.5 — 247 — — A 7.2 — 248 — — A 8.0 —249 — — A 7.7 — 267 — — A 10.7 — 268 — — A 10.0 — 269 — — A 12.2 — 270 —— A 12.3 — 271 — — A 9.3 — 272 — — A 12.7 — 273 — — A 12.7 — 274 — — A3.8 — 275 — — A 10.3 — 276 — — A 8.4 — 277 — — A 10.6 — 278 — — A 12.8 —279 — — A 11.4 — 280 — — A 7.9 — 281 — — A 11.5 — 282 — — A 8.6 — 283 —— A 8.4 — 284 — — A 12.2 — 290 — — A 11.4 — 291 — — A 9.7 — 292 — — A9.1 — 293 481.3 479.3 A 8.3 — 294 455.4 453.3 A 6.9 — 295 — — A 7.5 —296 — — A 8.9 — 298 353.4 — B 2.8 — 299 421.3 423.2 A 10.1 —

Example 6

[0151] Inhibition Assay

[0152] Compounds were assayed for the inhibition of ERK2 by aspectrophotometric coupled-enzyme assay (Fox et al (1998) Protein Sci 7,2249). In this assay, a fixed concentration of activated ERK2 (10 nM)was incubated with various concentrations of the compound in DMSO (2.5%)for 10 min. at 30° C. in 0.1 M HEPES buffer, pH 7.5, containing 10 mMMgCl2, 2.5 mM phosphoenolpyruvate, 200 μM NADH, 150 μg/mL pyruvatekinase, 50 μg/mL lactate dehydrogenase, and 200 μM erktide peptide. Thereaction was initiated by the addition of 65 μM ATP. The rate ofdecrease of absorbance at 340 nM was monitored. The IC₅₀ was evaluatedfrom the rate data as a function of inhibitor concentration.

[0153] Table 8 shows the results of the activity of selected compoundsof this invention in the ERK2 inhibition assay. The compound numberscorrespond to the compound numbers in Table 3. Compounds having anactivity designated as “A” provided a K_(i) value below 1 micromolar;compounds having an activity designated as “B” provided a K_(i) valuebetween 1 and 5 micromolar; and compounds having an activity designatedas “C” provided a K_(i) value greater than 5 micromolar. TABLE 8 ERK2Inhibitory Activity of Selected Compounds No. No No. II-A- ActivityII-A- Activity II-A- Activity 1 A 2 C 3 A 4 A 5 A 6 A 7 C 8 C 9 C 10 C11 C 12 C 13 A 14 C 16 C 17 C 18 A 19 A 20 A 21 C 22 A 23 A 24 A 25 C 26A 27 A 28 A 29 C 30 A 31 C 39 A 40 A 41 A 42 A 43 A 44 A 45 A 46 A 47 A48 A 49 A 50 A 51 A 52 A 53 A 54 A 55 A 56 A 57 A 58 A 59 A 60 A 61 A 62A 63 A 64 A I65 A 66 A 67 A 68 A 69 A 70 A 71 A 72 A 73 A 74 A 75 A 76 A77 A 78 A 79 A 80 A 81 A 82 A 83 A 84 A 85 A 86 A 87 A 88 A 89 A 90 A 91A 92 A 93 A 94 A 95 A 96 A 97 A 98 A 99 A 100 A 101 A 102 A 103 A 104 A105 A 106 A 107 A 108 A 109 A 110 A 111 A 112 A 113 A 114 A 115 A 116 B117 B 118 B 119 B 120 B 121 B 122 B 123 B 124 B 125 B 126 B 127 B 128 B129 B 130 B 131 B 132 B 133 B 134 B 135 B 136 B 137 B 138 B 139 B 140 B141 B 142 B 143 B 144 B 145 B 146 B 147 B 148 B 149 B 150 B 151 B 152 B153 B 154 B 155 B 156 B 157 B 158 B 159 B 160 B 161 C 162 C 163 C 164 C165 C 166 C 167 C 168 C 169 C 170 C 171 C 172 C 285 B 286 C 287 C 288 B289 C 290 B 291 C 292 C 293 C 294 C 295 C 296 C 297 C 298 C 299 C

Example 7

[0154] ERK Inhibition Cell Proliferation Assay

[0155] Compounds were assayed for the inhibition of ERK2 by a cellproliferation assay. In this assay, a complete media was prepared byadding 10% fetal bovine serum and penicillin/streptomycin solution toRPMI 1640 medium (JRH Biosciences). Colon cancer cells (HT-29 cell line)were added to each of 84 wells of a 96 well plate at a seeding densityof 10,000 cells/well/150 □L. The cells were allowed to attach to theplate by incubating at 37° C. for 2 hours. A solution of test compoundwas prepared in complete media by serial dilution to obtain thefollowing concentrations: 20 μM, 6.7 μM, 2.2 μM, 0.74 μM, 0.25 μM, and0.08 μM. The test compound solution (50 μL) was added to each of 72cell-containing wells. To the 12 remaining cell-containing wells, onlycomplete media (200 μL) was added to form a control group in order tomeasure maximal proliferation. To the remaining 12 empty wells, completemedia was added to form a vehicle control group in order to measurebackground. The plates were incubated at 37° C. for 3 days. A stocksolution of ³H-thymidine (1 mCi/mL, New England Nuclear, Boston, Mass.)was diluted to 20 μCi/mL in RPMI medium then 20 μL of this solution wasadded to each well. The plates were further incubated at 37° C. for 8hours then harvested and analyzed for ³H-thymidine uptake using a liquidscintillation counter.

[0156] Selected compounds of this invention that inhibit ERK in thecolon cell proliferation assay, with an IC₅₀ of less than 10 μM include:II-A-43, II-A-48, and II-A-45.

Example 8

[0157] The following example demonstrates a process for designing an ERKinhibitor of this invention:

[0158] Step 1) Choose a moiety containing a hydrogen bond acceptor asfrom Table 1, here pick pyrazole (Table 1 structure a).

[0159] This is Grp2.

[0160] Step 2) Confirm that the hydrogen bond acceptor from Grp2 iscapable of forming a satisfactory hydrogen bond with the backbone aminohydrogen of Met-108 of ERK.

[0161] In this case there is an HBD2 group that can also interactfavorably with the backbone carbonyl of Asp-106. Step 3) Choose a moietycontaining a hydrogen bond donor as from Table 2, here choose pyrrole(Table 2 structure aa)

[0162] This is Grp3.

[0163] Step 4) Confirm that the hydrogen bond donor from Grp3 is capableof forming a satisfactory hydrogen bond with the sidechain carbonyl ofGln-105.

[0164] Step 5) Choose a moiety capable of satisfying the distanceconstraints given for the relative positioning of Grp1 relative to Grp2and Grp3 while providing substantial attractive interactions with theenzyme environment, here pick benzene.

[0165] This is Grp1.

[0166] Step 6) Select a connectivity scheme for linking togethermoieties Grp1, Grp2, and Grp3 from structure types A, B, or C, asdescribed hereinabove.

[0167] Here we choose structure type A.

[0168] Step 7) Provide chemical links corresponding to the structuretype selected in Step 6. These links preferably contain the minimumnumber of bonds consistent with a chemically reasonable structure. Herewe pick a valence bond to connect Grp1 to Grp2, and a valence bond toconnect Grp 2 to Grp3.

[0169] Step 8) Minimize the entire constructed molecule in the contextof the active site in order to provide additional R-groups that may becapable of providing further binding to the enzyme (e.g. satisfyingunpaired hydrogen bond donors or acceptors or satisfying hydrophobicinteractions). Here we connect a —C(═O)NH—CH₂—Ph to the 2-position ofthe pyrrole ring in order to provide a hydrogen bond acceptor (C═O) forthe sidechain NH₃ of Lys-52 and a hydrophobic group (Ph) for theC(alpha) through C(delta) of Lys-52 and surrounding hydrophobic groups,resulting in compound II-A-5 below.

[0170] While we have described a number of embodiments of thisinvention, it is apparent that our basic examples may be altered toprovide other embodiments which utilize the compounds and methods ofthis invention. Therefore, it will be appreciated that the scope of thisinvention is to be defined by the appended claims rather than by thespecific embodiments which have been represented by way of example.

We claim:
 1. A method of inhibiting protein kinase activity in abiological sample, wherein said protein kinase is ERK, comprising thestep of contacting said sample with a compound comprising Grp1, Grp2,and Grp3, wherein: Grp 1 is an optionally substituted aryl or aliphaticgroup; Grp 2 is a heteroaromatic ring comprising one to three nitrogens,and a hydrogen bond acceptor HBA2, wherein HBA2 is optionally bonded toa hydrogen bond donor HBD2; and Grp3 is a heteroaromatic ring comprisinga hydrogen bond donor HBD1; wherein said compound optionally comprises ahydrogen bond acceptor HBA1; and wherein Grp1 is within about 2.5-10.0 Åof Grp2; Grp2 is within about 4.0-8.0 Å of Grp3; Grp3 is within about5.0-12.0 Å of Grp1; HBA2 is within about 6.5-11.0 Å of Grp1; HBD1 iswithin about 6.5-8.5 Å of Grp2; HBD1 is within about 3.5-5.5 Å of HBA1;and HBA1 is within about 6.7-14.0 Å of HBD2.
 2. The method according toclaim 1 wherein the compound comprises hydrogen bond acceptor HBA1. 3.The method according to claim 2 wherein the hydrogen bond acceptor HBA2is bonded to a hydrogen bond donor HBD2.
 4. The method according to anyof claims 1 to 3, wherein: Grp3 is within about 3.9-8.0 Å of Grp2; Grp2is within about 5.5-6.6 Å of Grp3; Grp3 is within about 6.0-10.0 Å ofGrp1; HBA2 is within about 6.5-11.0 Å of Grp1; HBD1 is within about7.2-8.2 Å of Grp2; HBD1 is within about 3.9-4.9 Å of HBA1; and HBA1 iswithin about 7.7-11.7 Å or 11.6-13.6 Å of HBD2.
 5. The method accordingto claim 4, wherein: Grp1 is within about 5.7-6.8 Å of Grp2; Grp2 iswithin about 5.5-6.6 Å of Grp3; Grp3 is within about 7.2-8.2 Å of Grp1;HBA2 is within about 6.5-11.0 Å of Grp1; HBD1 is within about 7.2-8.2 Åof Grp2; HBD1 is within about 3.9-4.9 Å of HBA1; and HBA1 is withinabout 10.2-11.2 Å or 12.1-13.1 Å of HBD2.
 6. The method according toclaim 5 wherein Grp2 is a ring selected from Table
 1. 7. The methodaccording to claim 6 wherein Grp3 is a ring selected from Table
 2. 8. Acomposition comprising a compound in an amount sufficient to detectablyinhibit protein kinase activity, said protein kinase is ERK; and apharmaceutically acceptable carrier, wherein said compound comprisesGrp1, Grp2 and Grp3, wherein: Grp 1 is an optionally substituted aryl oraliphatic group; Grp 2 is a heteroaromatic ring comprising one to threenitrogens, and a hydrogen bond acceptor HBA2, wherein HBA2 is optionallybonded to a hydrogen bond donor HBD2; and Grp3 is a heteroaromatic ringcomprising a hydrogen bond donor HBD1; wherein said compound optionallycomprises a hydrogen bond acceptor HBA1; and wherein Grp1 is withinabout 2.5-10.0 Å of Grp2; Grp2 is within about 4.0-8.0 Å of Grp3; Grp3is within about 5.0-12.0 Å of Grp1; HBA2 is within about 6.5-11.0 Å ofGrp1; HBD1 is within about 6.5-8.5 Å of Grp2; HBD1 is within about3.5-5.5 Å of HBA1; and HBA1 is within about 6.7-14.0 Å of HBD2.
 9. Thecomposition according to claim 8 wherein the compound comprises hydrogenbond acceptor HBA1.
 10. The composition according to claim 9 wherein thehydrogen bond acceptor HBA2 is bonded to a hydrogen bond donor HBD2. 11.The composition according to claim 10, wherein: Grp1 is within about3.9-8.0 Å of Grp2; Grp2 is within about 5.5-6.6 Å of Grp3; Grp3 iswithin about 6.0-10.0 Å of Grp1; HBA2 is within about 6.5-11.0 Å ofGrp1; HBD1 is within about 7.2-8.2 Å of Grp2; HBD1 is within about3.9-4.9 Å of HBA1; and HBA1 is within about 7.7-11.7 Å or 11.6-13.6 Å ofHBD2.
 12. The composition according to claim 11, wherein: Grp1 is withinabout 5.7-6.8 Å of Grp2; Grp2 is within about 5.5-6.6 Å of Grp3; Grp3 iswithin about 7.2-8.2 Å of Grp1; HBA2 is within about 6.5-11.0 Å of Grp1;HBD1 is within about 7.2-8.2 Å of Grp2; HBD1 is within about 3.9-4.9 Åof HBA1; and HBA1 is within about 10.2-11.2 Å or 12.1-13.1 Å of HBD2.13. The composition according to claim 12 wherein Grp2 is a ringselected from Table
 1. 14. The composition according to claim 13 whereinGrp3 is a ring selected from Table
 2. 15. The composition according toclaim 14 wherein said compound is formulated in a pharmaceuticallyacceptable manner for administration to a patient.
 16. The compositionaccording to claim 15 further comprising a therapeutic agent, either aspart of a multiple dosage form together with said compound or as aseparate dosage form.
 17. A method for treating a disease state in apatient that is alleviated by treatment with a protein kinase inhibitor,wherein said protein kinase is ERK, comprising administering to saidpatient in need of such a treatment a therapeutically effective amountof a composition according to any of claims 8 to
 15. 18. The methodaccording to claim 17, comprising the additional step of administeringto said patient a therapeutic agent either as part of a multiple dosageform together with said compound or as a separate dosage form.
 19. Themethod according to claim 19 wherein said disease state is selected fromcancer, stroke, diabetes, hepatomegaly, cardiovascular disease,Alzheimer's disease, cystic fibrosis, viral disease, autoimmunediseases, atherosclerosis, restenosis, psoriasis, allergic disorders,inflammation, neurological disorders, a hormone-related disease,conditions associated with organ transplantation, immunodeficiencydisorders, destructive bone disorders, proliferative disorders,infectious diseases, conditions associated with cell death,thrombin-induced platelet aggregation, chronic myelogenous leukemia(CML), liver disease, pathologic immune conditions involving T cellactivation, or CNS disorders.
 20. A method of treating a disease statein a patient, said disease state selected from cancer, stroke, diabetes,hepatomegaly, cardiovascular disease, Alzheimer's disease, cysticfibrosis, viral disease, autoimmune diseases, atherosclerosis,restenosis, psoriasis, allergic reactions, inflammation, neurologicaldisorders or a hormone-related disease, which method comprisesadministering to a patient in need of such a treatment a therapeuticallyeffective amount of a composition according to any of claims 8 to 15.21. The method according to claim 20, comprising the additional step ofadministering to said patient a therapeutic agent either as part of amultiple dosage form together with said compound or as a separate dosageform.
 22. The method according to claim 20 wherein the disease state iscancer.
 23. The method according to claim 20 wherein the disease stateis a cardiovascular disease.
 24. A method of designing an ERK inhibitor,comprising the steps of: a) selecting a Grp2 moiety containing ahydrogen bond acceptor HBA2, wherein said HBA2 forms a hydrogen bondwith the backbone amino hydrogen of Met-108; b) selecting a Grp3 moietyand a means for attachment to Grp2, wherein Grp3 so attached is withinthe requisite distances to Grp2 and HBA2, and HBD1 forms a hydrogen bondwith the sidechain carbonyl of Gln-105; c) selecting a Grp1 moiety and ameans for attachment to the fragment formed by steps a) through c); d)selecting a structure type for connecting Grp1, Grp2, and Grp3; e)connecting Grp1, Grp2, and Grp3 according to the structure type selectedin step f); f) minimizing the constructed molecule within the proteinkinase acitve site; g) optionally selecting an HBA1 group and connectingsaid HBA1 group to the molecule minimized in step h);
 25. A compositionfor coating an implantable device comprising a compound and a carriersuitable for coating said implantable device; wherein said compoundcomprises Grp1, Grp2 and Grp3, wherein: Grp 1 is an optionallysubstituted aryl or aliphatic group; Grp 2 is a heteroaromatic ringcomprising one to three nitrogens, and a hydrogen bond acceptor HBA2,wherein HBA2 is optionally bonded to a hydrogen bond donor HBD2; andGrp3 is a heteroaromatic ring comprising a hydrogen bond donor HBD1;wherein said compound optionally comprises a hydrogen bond acceptorHBA1; and wherein Grp1 is within about 2.5-10.0 Å of Grp2; Grp2 iswithin about 4.0-8.0 Å of Grp3; Grp3 is within about 5.0-12.0 Å of Grp1;HBA2 is within about 6.5-11.0 Å of Grp1; HBD1 is within about 6.5-8.5 Åof Grp2; HBD1 is within about 3.5-5.5 Å of HBA1; and HBA1 is withinabout 6.7-14.0 Å of HBD2.
 26. An implantable device coated with acomposition according to claim 25.